Automated processing systems and methods of thermally processing microscope slides

ABSTRACT

Methods and apparatus that enable drying and curing a plurality of specimens carried by a plurality of microscope slides. Slide carriers are positioned at a first position while the slide carrier holds the microscope slides. Each of the specimens can be carried by one of the microscope slides. The slide carrier can be robotically moved to move the slide carrier into a circulation loop defined by a heater apparatus. The specimens and/or microscope slides can be convectively heated while the slide carrier is located in the circulation loop.

CROSS-REFERENCE TO RELATED APPLICATIONS

This patent application is a continuation of International PatentApplication No. PCT/EP2014/076894 filed Dec. 8, 2014, which claimspriority to and the benefit of U.S. Provisional Patent Application No.61/916,107 filed Dec. 13, 2013. Each patent application is incorporatedherein by reference as if set forth in its entirety.

TECHNICAL FIELD

The present technology is generally related to automated histologicalprocessing of biological specimens (e.g., tissue samples), such assystems, devices, methods, and compositions that enhance the quality,precision, efficiency and/or other aspects of this processing.

BACKGROUND

A wide variety of techniques may be used to analyze biologicalspecimens. Examples of analysis techniques useful in this contextinclude microscopy, microarray analysis (e.g., protein and nucleic acidmicroarray analysis), and mass spectrometry. Preparing specimens forthese and other types of analysis typically includes contacting thespecimens with a series of processing liquids. Some of these processingliquids (e.g., staining reagents and counterstaining reagents) may addcolor and contrast or otherwise change the visual characteristics ofinvisible or poorly visible specimen components (e.g., at least sometypes of cells and intracellular structures). Other processing liquids(e.g., deparaffinizing liquids) may be used to achieve other processingobjectives. If a specimen is treated with multiple processing liquids,both the application and the subsequent removal of each processingliquid can be important for producing specimens suitable for analysis.In some cases, treating specimens with multiple processing liquidsincludes manually applying the processing liquids to microscope slidesrespectively carrying the specimens. This approach to processingspecimens tends to be relatively labor intensive and imprecise.

“Dip and dunk” automated machines can be used as an alternative tomanual specimen processing. These machines automatically processspecimens by submerging racks of specimen-bearing slides in open bathsof processing liquids. Unfortunately, operation of dip and dunk machinesinevitably causes carryover of processing liquids from one bath toanother. Over time, this carryover leads to the degradation of theprocessing liquids. Furthermore, when specimens are immersed in a sharedbath, there is a potential for cross-contamination. For example, cellsmay slough off a specimen on one slide and be transported within ashared bath onto another slide, even on a slide processed much later(e.g., if the cells remain suspended in the bath). This form ofcontamination can adversely affect the accuracy of certain types ofspecimen analysis. To mitigate this issue and to address degradation ofprocessing liquids due to carryover, baths of processing liquids in dipand dunk machines typically need to be replaced frequently. Accordingly,these machines tend to consume relatively large volumes of processingliquids, which increases the economic and environmental costs associatedwith operating these machines. Open baths of processing liquids are alsoprone to evaporative losses and oxidative degradation of someprocessing-liquid components. Oxidation of certain components ofstaining reagents, for example, can alter the staining performance ofthese components and thereby adversely affect the precision of stainingoperations.

Some example of conventional histological processing machines that avoidcertain disadvantages of dip and dunk machines are known. For example,U.S. Pat. No. 6,387,326 (the '326 patent) to Edwards et al. describes anapparatus for delivering fresh processing liquids directly ontoindividual slides. The slides are expelled one at a time from a slidestorage device onto a conveyor belt. Specimens carried by the slides areindividually treated at various stations as the slides move along theconveyor belt. Among other drawbacks, the apparatus described in the'326 patent and similar machines tend to have throughput limitationsthat make them unsuitable for primary staining applications, such ashematoxylin and eosin (H&E) staining applications. A typical laboratorythat performs primary staining, for example, may process hundreds oreven thousands of specimens per day. Using the apparatus described inthe '326 patent and similar machines for this processing would beunacceptably slow. Furthermore, these machines do not allow for controlover staining characteristics. Such control can be important in primarystaining applications.

Overview of Technology

At least some embodiments are an automated system configured to performone or more slide processing operations on slides bearing biologicalsamples. The system can provide high sample throughput while alsominimizing or limiting the potential for cross-contamination of slides.The automated systems can include features that facilitate consistency,controllability of processing time, and/or processing temperature.

At least some embodiments are a method for drying a plurality ofspecimens carried by a plurality of microscope slides. The methodincludes positioning a slide carrier at a first position while the slidecarrier holds the microscope slides. Each of the specimens can becarried by one of the microscope slides. The slide carrier can berobotically moved to move the slide carrier into a circulation loopdefined by a heater apparatus. The specimens and/or microscope slidescan be heated while the slide carrier is located in the circulationloop. In certain embodiments, the specimens and/or microscope slides canbe convectively, conductively, and/or radiantly heated.

In some embodiments, a heater apparatus for heating a plurality ofspecimens carried by a plurality of microscope slides includes ahousing, a blower, and a door assembly. The housing can at leastpartially define a circulation loop. The blower can be positioned toproduce a fluid flow along the circulation loop. The door assembly ismoveable between a first position and a second position. In someembodiments, the apparatus includes a heat source configured to heat thefluid flow such that the specimens are convectively heated by the fluidflow when the door assembly holds a slide carrier along the circulationloop.

The apparatus, in some embodiments, can be configured to provideconductive and/or radiant heating. Conductive heating can be providedvia a plate with a resistive heater. One or more lamps can provideradiant heating. The apparatus can controllably increase or decrease thetemperature of the specimens. In some embodiments, when in the firstposition, the door assembly can be configured to receive the slidecarrier that carries the microscope slides. When in the second position,the door assembly can be configured to hold the slide carrier at avertically-oriented position along the circulation loop. The doorassembly can also be moved to other positions.

In some embodiments, a method for thermally processing coverslips isprovided. One or more specimens can be covered by a coverslip andcarried by one of a plurality of microscope slides. The method includespositioning a slide carrier at a first position while the slide carrierholds the microscope slides. The slide carrier can be roboticallypositioned at a second position within a circulation loop defined by aheater apparatus. In some embodiments, convective heating is used toheat the coverslips and/or microscope slides positioned within thecirculation loop. Conductive and/or radiant heating can also be used.For example, convective heating/cooling can be used for one or moreperiods of time and radiant heating can be used for one or more periodsof time.

At least some embodiments can be a method for processing a specimencarried by a slide within an automated histological system. The methodincludes automatically dispensing a first liquid so as to form a firstpuddle on the slide. The first puddle has a shape maintained at leastpartially by surface tension and can be one of a staining reagent and acounterstaining reagent. The specimen is stained with the first liquidwhile the specimen is in contact with the first puddle. At least aportion of the first puddle is removed from the specimen so as to atleast partially uncover the specimen a first time. The specimen iscontacted with an intermediate fluid after at least partially uncoveringthe specimen the first time. The specimen is at least partiallyuncovered a second time after contacting the intermediate fluid and thespecimen. A second liquid is automatically dispensed so as to form asecond puddle on the slide. The second puddle has a shape maintained atleast partially by surface tension, and the second liquid can be theother of the staining reagent and the counterstaining reagent. Thespecimen can be stained by the second liquid while the specimen is incontact with the second puddle, for example, after at least partiallyuncovering the specimen the second time.

In some embodiments, a method for processing specimens carried by slideswithin an automated histological system includes dispensing a liquid soas to form a first puddle on a first slide. The liquid can be one of astaining reagent and a counterstaining reagent. Liquid can be dispensedso as to form a second puddle on a second slide. The first and secondspecimens can be stained (e.g., non-immunohistochemically stained) whilethe first and second specimens are in contact with the first and secondpuddles, respectively. At least a portion of the first puddle is removedfrom the first specimen so as to at least partially uncover the firstspecimen without contacting the first puddle with a solid structureand/or displacing the first puddle with a liquid. At least a portion ofthe second puddle can be removed from the second specimen so as to atleast partially uncover the second specimen without contacting thesecond puddle with a solid structure or displacing the second puddlewith a liquid. In some embodiments, the first and second puddles arefreestanding puddles.

At least some embodiments are a method that includes delivering a liquidfrom a fluid dispense mechanism at an anti-splatter fluid exit speed.The liquid flows at the anti-splatter fluid exit speed and is directedtoward a microscope slide (e.g., an upper surface of the slide) suchthat the microscope slide carries a collected volume of the liquid. Theliquid can be at least partially supported on the slide by, for example,surface tension. In some embodiments, the anti-splatter fluid exit speedis less than a splattering fluid exit speed at which the directed liquidwould tend to cause at least a portion of the collected volume tosplatter from the upper surface. In some embodiments, the anti-splatterfluid exit speed is greater than a trampoline fluid exit speed at whichat least a portion of the directed liquid would tend to bounce off asurface of the collected volume of liquid.

In some embodiments, a method for processing one or more microscopeslides includes delivering a liquid at an anti-splatter fluid flow ratethat is less than a splattering fluid flow rate at which the directedliquid would tend to cause at least a portion of the collected volume tonot stay on the slide. For example, the anti-splatter fluid flow ratecan be sufficiently low to prevent appreciable splattering of thecollected liquid. In some embodiments, the anti-splatter flow rate isgreater than a trampoline flow rate at which at least a portion of thedirected liquid would tend to bounce off a surface of the collectedvolume of liquid. The anti-splatter flow rate can be selected based oncharacteristics of the liquid.

In yet other embodiments, a method for processing a specimen on an uppersurface of a microscope slide includes moving the microscope slide to aprocessing position. A liquid barrier material can be dispensed onto themicroscope slide at the processing position to form a barrier comprisedof the barrier material along at least a portion of a label of themicroscope slide. A liquid (e.g., reagent) can be delivered onto themicroscope slide such that the liquid contacts the specimen while thebarrier covers at least the portion of the label. In some embodiments,the microscope slide can be robotically moved to the processing positionusing a an automated mechanism, such as a transport mechanism.

In yet further embodiments, a method for processing a specimen on amicroscope slide includes dispensing reagent from outlets of a fluiddispense mechanism aligned with a width of an upper surface of themicroscope slide. The width of the upper surface can be substantiallyperpendicular to a longitudinal axis of the microscope slide. Theoutlets can be moved in a direction substantially parallel to thelongitudinal axis of the slide to distribute the reagent within amounting area of the upper surface so as to form a layer of the reagentthat contacts a specimen located at the mounting area.

At least some embodiments are a system for processing a specimen on amicroscope slide includes a transporter device, an automated slideprocessing module, and a dispenser assembly. The automated slideprocessing module can be positioned to receive a slide carrier from thetransporter device and can include a dispenser assembly movable along amicroscope slide held by the slide carrier when the slide carrier islocated within a holding chamber. The dispenser assembly includes aplurality of outlets configured to be aligned with a width of an uppersurface of the microscope slide such that the outlets apply a reagentacross most or all of the width of the upper surface.

In some embodiments, a system comprises a transporter device and astainer module configured to receive a slide carrier from thetransporter device. In certain embodiments, the stainer module includesone or more fluid lines and a head assembly movable to dispense reagentalong a slide carried by the slide carrier. The head assembly can becoupled to the fluid lines and can be configured to dispense reagentfrom one or all of the fluid lines. In one embodiment, a manifold of thehead assembly includes a distribution chamber, a plurality of inletsopening into the distribution chamber, and a plurality of outlets fromthe distribution chamber. The fluid can be delivered through themanifold and dispensed from the head assembly.

In yet further embodiments, a microscope slide processing systemcomprises a transporter device and a stainer module configured toreceive a slide carrier from the transporter device. The stainer modulecan include a plurality of manifolds and a plurality of nozzles in fluidcommunication with the manifolds. In some embodiments, the stainermodule includes a plurality of first fluid lines, a plurality of secondfluid lines, and a dispenser head movable relative the slide carrier, ifany, positioned in the stainer module. The dispenser head can comprise aplurality of first nozzles, a first manifold configured to distributefluid from each of the first fluid lines to the first nozzles, aplurality of second nozzles, and a second manifold configured todistribute fluid from each of the second fluid lines to the secondnozzles. The dispenser head can include additional manifolds and/ornozzles to distribute liquid from any number of fluid lines.

At least some embodiments are an automated slide processing apparatusfor staining a specimen on a microscope slide located within the slideprocessing apparatus. The slide processing apparatus includes a liquidremoval device, a gas knife, and a suction element. The liquid removaldevice is movable relative to the slide. In some embodiments, the gasknife generates a gas curtain and a low pressure region to facilitateliquid removal. In some embodiments, the gas knife is configured togenerate a gas curtain that tends to collect liquid on an upper surfaceof the slide at a collection zone at least partially defined by the gascurtain as the liquid removal device moves relative to the slide. Asuction element is positioned to remove liquid collected at thecollection zone from the upper surface as the liquid removal devicemoves relative to the slide.

In some embodiments, a slide processing apparatus for staining aspecimen on a microscope slide located within the slide processingapparatus comprises a fluid removal device movable relative to theslide. The fluid removal device includes a fluid knife configured tooutput one or more gas flows to urge a volume of liquid on an uppersurface of the slide toward a collection zone on the upper surface. Thecollection zone can be at least partially defined by the one or more gasflows. In certain embodiments, the collection zone is a centralcollection zone. In other embodiments, the collection zone is at otherlocations along the slide.

In another embodiment, a slide processing apparatus comprises a suctionelement and a fluid knife movable relative to a microscope slide tocaptivate at least a portion of a volume of liquid on the slide. Thesuction element and the gas knife are configured to cooperate to drawmost or all of the volume of liquid into the suction element. In someembodiments, the slide processing apparatus includes a plurality ofsuction elements to draw in liquid at different locations.

In yet another embodiment, a method for processing a specimen on amicroscope slide includes applying a liquid onto the slide to cover thespecimen with the liquid. A stream of fluid is delivered toward an uppersurface of the slide to move the applied liquid along the upper surfacewhile confining the applied liquid such that the confined liquid isincreasingly spaced apart from longitudinal edges of the slide. Theconfined liquid is removed from the upper surface of the slide.

In some embodiments, a method for processing a specimen on a microscopeslide includes applying a liquid onto the slide and directing anon-planar or multiplanar gas curtain toward an upper surface of theslide. A vertex section of the gas curtain can be moved along a centralregion of the upper surface and toward an end of the slide so as to urgethe applied liquid toward the central region of the slide. In otherembodiments, the vertex section of the gas curtain can be moved alongother regions of the upper surface.

In particular embodiments, a method for processing a specimen on amicroscope slide includes delivering the slide into a stainer module.Liquid is applied onto the slide to contact the specimen with theliquid. The liquid is blown along and removed from an upper surface ofthe slide. The slide can then be removed from the stainer module. Insome embodiments, the slides are robotically delivered into and/orremoved from the stainer module.

At least some embodiments are a method that includes moving a headassembly of a stainer module relative to a first microscope slidepositioned at a processing zone within the stainer module so as to applyone or more reagents onto the first microscope slide. After applying theone or more reagents onto the first microscope slide, the firstmicroscope slide is moved away from the processing zone and a secondmicroscope slide is moved to the processing zone. The head assembly ismoved relative to the second microscope slide while the secondmicroscope slide is positioned at the processing zone so as to apply oneor more reagents onto the second microscope slide.

In some embodiments, a method for processing a plurality of microscopeslides carrying specimens using a stainer module includes delivering aslide carrier tray carrying the microscope slides into the stainermodule. The stainer module includes a movable dispenser apparatus havinghead assemblies. At least one of the microscope slides is processed bydelivering one or more liquids from the dispenser assembly while theslide carrier tray obstructs a first set of vertical delivery paths froma first set of the head assemblies and obstructs a second set ofvertical delivery paths from a second set of the head assemblies. Theslide carrier tray can be moved to a purge position to unobstruct thefirst set of vertical delivery paths such that the collection pancollects liquid outputted by the first set of the head assemblies. Theslide carrier tray can be moved to a second position to unobstruct thesecond set of vertical delivery paths such that the collection pancollects liquid outputted by the second set of the head assemblies. Thefirst set can be different from the second set.

In additional embodiments, an apparatus for processing a plurality ofmicroscope slides includes at least one stainer module. The stainermodule can include a tray holder and a head assembly. The tray holdercan be configured to receive and hold a tray carrying a first microscopeslide and a second microscope slide in a chamber of the stainer module.The head assembly is movable relative to a processing zone in thestainer module so as to deliver one or more liquids outputted from thehead assembly along the first microscope slide positioned at theprocessing zone. In some embodiments, the tray holder is movable totransport the first microscope slide away from the processing zone andto transport the second microscope slide to the processing zone afterdelivering the one or more liquids onto the first microscope slide.

In yet additional embodiments, an apparatus for processing a pluralityof microscope slides comprises a stainer module including fluid lines, atray holder, and a head assembly. The tray holder is configured toreceive and hold a tray carrying a first microscope slide and a secondmicroscope slide in the stainer module. The head assembly includes adispenser head and one or more valves mounted on the dispenser head. Thevalves can control which fluid from the plurality of fluid lines flowsthrough and out of the head. The dispenser head can carry the valves andis movable relative to tray holder so as to deliver one or more fluidsoutputted from the dispenser head along the microscope slides.

At least some embodiments are directed to a method for processingspecimens carried by slides within an automated histological stainingsystem. The method includes moving a slide carrier toward and into atemperature-controlled internal environment of a stainer within thesystem. The slide carrier carries a first slide and a second slide, andthe first and second slides can carry a first specimen and a secondspecimen, respectively. The first and second specimens are stained withat least one of a staining reagent and a counterstaining reagent whilethe first and second slides are within the internal environment andwhile an average temperature of the internal environment is greater thanambient temperature. The slide carrier can be moved out of the internalenvironment after staining one or both specimens.

In some embodiments, an automated histological staining system comprisesa main housing and a stainer. The stainer includes a stainer housingdefining an internal environment of the stainer, one or more heatersconfigured to internally heat the stainer, and a transporter. Thetransporter can be configured to move a slide carrier robotically withinthe main housing toward the stainer. In one embodiment, the transportermoves the slide carrier between multiple modules in the main housing.

At least some embodiments are directed to a method for processingspecimens in an automated histological staining system. The methodcomprises robotically moving a slide carrier into a stainer of thesystem. The slide carrier carries slides which respectively carry thespecimens, and the specimens are at least partially embedded inparaffin. Liquids are automatically dispensed onto the slides accordingto a predetermined recipe for at least deparaffinizing, staining, andcounterstaining the specimens. The slide carrier can be roboticallymoved out of the stainer after automatically dispensing the liquids. Insome embodiments, a total of all liquid dispensed onto the slides aftermoving the slide carrier into the stainer and before moving the slidecarrier out of the stainer has a greater volumetric concentration ofpolyol than of monohydric alcohol.

In one embodiment, a method for processing specimens within an automatedhistological staining system comprises contacting the specimens with astaining reagent. The specimens can be contacted by a washing liquid toat least partially remove the staining reagent from the specimens. Thespecimens can be contacted with a counterstaining reagent aftercontacting the specimens and the washing liquid. The specimens can becontacted with the washing liquid to differentiate counterstaining ofthe specimens after contacting the specimens and the counterstainingreagent. In some embodiments, one or more of the staining reagent,washing liquid, and/or counterstaining reagent has a greater volumetricconcentrations of polyol than of monohydric alcohol. In one embodiment,the staining reagent, the washing liquid, and the counterstainingreagent each have greater volumetric concentrations of polyol than ofmonohydric alcohol.

BRIEF DESCRIPTION OF THE DRAWINGS

Many aspects of the present disclosure can be better understood withreference to the following drawings. The relative dimensions in thedrawings may be to scale with respect to some embodiments. With respectto other embodiments, the drawings may not be to scale. For ease ofreference, throughout this disclosure identical reference numbers may beused to identify identical or at least generally similar or analogouscomponents or features.

FIG. 1 is a front elevation view of an automated slide processing systemin accordance with an embodiment of the present technology.

FIG. 2 is a front elevation view of the automated slide processingsystem of FIG. 1 showing internal components of the system.

FIG. 3 is a cross-sectional perspective view of a dryer apparatusheating specimen-bearing microscope slides in accordance with anembodiment of the present technology.

FIG. 4A is a side elevation view of a dryer apparatus having a door inan open configuration in accordance with an embodiment of the presenttechnology.

FIG. 4B is an enlarged perspective view of a door assembly of the dryerapparatus of FIG. 4A.

FIG. 5 is a perspective view of the dryer apparatus of FIG. 4A in anopen configuration holding a slide carrier.

FIG. 6A is an enlarged cross-sectional side elevation view of the dryerapparatus of FIG. 4A in a closed configuration supporting the slidecarrier of FIG. 5 in accordance with an embodiment of the presenttechnology.

FIG. 6B is an enlarged cross-sectional side elevation view of a portionof FIG. 6B.

FIG. 7 is an enlarged cross-sectional side elevation view of a portionof a dryer apparatus door assembly and a slide carrier in asubstantially vertical position in accordance with another embodiment ofthe present technology.

FIG. 8 is a perspective view of the dryer apparatus of FIG. 4A in anopen configuration holding a slide carrier in accordance with anembodiment of the present technology.

FIG. 9 is a cross-sectional side elevation view of the dryer apparatusof FIG. 4A in a closed configuration without a slide carrier inaccordance with an embodiment of the present technology.

FIG. 10 is a perspective view of a curing oven in a closed configurationin accordance with an embodiment of the present technology.

FIG. 11 is a perspective view of the curing oven of FIG. 10 in an openconfiguration in accordance with an embodiment of the presenttechnology.

FIG. 12 is a perspective view of the curing oven with a door assemblysupporting a slide carrier holding microscope slides with coverslips inaccordance with an embodiment of the present technology.

FIG. 13 is a cross-sectional side elevation view of a curing oven in aclosed configuration in accordance with an embodiment of the presenttechnology.

FIG. 14 is a perspective view of a curing oven in an open configurationholding a slide carrier with cured coverslipped slides in accordancewith an embodiment of the present technology.

FIG. 15 is an isometric view of a stainer module in an openconfiguration in accordance with an embodiment of the presenttechnology.

FIG. 16 is an isometric view of the stainer module holding a tray.

FIG. 17 is a bottom view of the stainer module holding the tray.

FIG. 18 is a bottom view of the stainer module in a closed configurationin accordance with an embodiment of the present technology.

FIG. 19 is an isometric view of the stainer module of FIG. 15 ready toprocess specimen-bearing slides positioned underneath a dispenserapparatus.

FIG. 20 is a top plan view of the stainer module of FIG. 15.

FIG. 21 is a cross-sectional side elevation view of the stainer moduletaken along line 21-21 of FIG. 20.

FIGS. 22A and 22B are detailed elevation views of head assembliesprocessing specimen-bearing microscope slides.

FIG. 23 is an isometric view of a tray holding microscope slides inaccordance with an embodiment of the present technology.

FIGS. 24-26 are perspective views of stages of applying substances tomicroscope slides.

FIG. 27 is a top plan view of lower components of the stainer modulealong line 27-27 of FIG. 21.

FIG. 28 is a cross-sectional side elevation view of a liquid collectortaken along line 28-28 of FIG. 27.

FIGS. 29A-31B are top plan and side elevation views show stages of apurge/prime process in accordance with an embodiment of the presenttechnology.

FIG. 32 is an isometric view of a dispenser apparatus in accordance withan embodiment of the present technology.

FIG. 33 is a side elevation view of a head assembly dispensing liquidonto a microscope slide in accordance with an embodiment of the presenttechnology.

FIG. 33A is a detailed view of a nozzle of the head assembly of FIG. 33.

FIG. 34 is an isometric view of a head assembly in accordance with anembodiment of the present technology.

FIG. 35 is a bottom view of the head assembly of FIG. 34 and amicroscope slide.

FIG. 36 is a side elevation view of the head assembly dispensing liquidonto a label of a microscope slide in accordance with an embodiment ofthe present technology.

FIG. 36A is a detailed view of a nozzle directing a stream of liquidtoward the label.

FIG. 37 is a side elevation view of the head assembly dispensing liquidonto a mounting area of the microscope slide.

FIG. 38 is a side elevation view of the head assembly dispensing liquidonto an end of the microscope slide.

FIGS. 39, 40, and 41 are isometric, side, and front views, respectively,of the head assembly in accordance with an embodiment of the presenttechnology.

FIG. 42A is a cross-sectional perspective view of the head assemblytaken along line 42A-42A of FIG. 41.

FIG. 42B is a detailed view of manifolds of the head assembly of FIG.42A.

FIG. 43 is a cross-sectional elevation view of the head assembly takenalong line 43-43 of FIG. 40.

FIG. 44A is a cross-sectional perspective view of the head assemblytaken along line 44A-44A of FIG. 41.

FIG. 44B is a detailed view of manifolds of the head assembly of FIG.44A.

FIG. 45 is a cross-sectional elevation view of the head assembly takenalong line 45-45 of FIG. 40.

FIGS. 46A-46F are cross-sectional elevation views of the head assemblytaken along line 46-46 of FIG. 40.

FIG. 47 is an isometric cross-sectional view of the head assembly takenalong line 47-47 of FIG. 40.

FIGS. 48A-C are cross-sectional views of the head assembly taken alongline 48-48 of FIG. 41.

FIG. 49 is an isometric view of a head assembly in accordance with anembodiment of the present technology.

FIG. 50 is a top plan view of the head assembly of FIG. 49.

FIG. 51 is an isometric view of a dispenser head in accordance with anembodiment of the present technology.

FIG. 52 is a cross-sectional perspective view of the dispenser headtaken along line 52-52 of FIG. 50.

FIG. 53 is an isometric view of a liquid distributor device inaccordance with an embodiment of the present technology.

FIG. 54 is a cross-sectional elevation view of a nozzle apparatus inaccordance with an embodiment of the present technology.

FIG. 55 is an isometric view of a dispenser apparatus in accordance withan embodiment of the present technology.

FIGS. 56-58 are side elevation views illustrating stages of a liquidremoval process in accordance with an embodiment of the presenttechnology.

FIGS. 59-61 are isometric, front, and bottom views, respectively, of ahead assembly in accordance with an embodiment of the presenttechnology.

FIG. 62 is a partial cross-sectional side view of a liquid removaldevice positioned above a microscope slide.

FIG. 63 is a partial cross-sectional side view of the liquid removaldevice sucking liquid from the slide.

FIG. 64A is an isometric view of a liquid removal device producing a gascurtain positioned along a microscope slide in accordance with anembodiment of the present technology.

FIG. 64B is a top plan view of the gas curtain and the slide of FIG.64A.

FIG. 65A is an isometric view of the liquid removal device collectingliquid using the gas curtain.

FIG. 65B is a top plan view of the gas curtain and slide of FIG. 65A.

FIG. 66A is an isometric view of the liquid removal device captivatingliquid at an end of the slide.

FIG. 66B is a top plan view of the gas curtain and slide of FIG. 66A.

FIGS. 67-70 are side elevation views illustrating stages of removing anddispensing liquids in accordance with an embodiment of the presenttechnology.

FIG. 71 is an isometric view of a liquid removal device with a lineargas knife in accordance with an embodiment of the present technology.

FIG. 72 is an isometric view of the liquid removal device of FIG. 71collecting liquid along the slide.

FIG. 73 is an isometric view of the liquid removal device of FIG. 71removing liquid captivated at a corner of the slide.

FIG. 74 is a bottom view of a liquid removal device with a gas knifehaving elongated slots in accordance with an embodiment of the presenttechnology.

FIG. 75 is a bottom view of a liquid removal device with two gas knivesin accordance with an embodiment of the present technology.

FIG. 76 is an isometric view of two gas knives in accordance with anembodiment of the present technology.

FIGS. 77 and 78 are side elevation views of two gas knives captivatingliquid on a microscope slide.

FIG. 79 is an isometric view of a stainer configured in accordance withan embodiment of the present technology.

FIG. 80 is a cross-sectional side elevation view taken along the line80-80 in FIG. 79 showing an internal environment of the stainer.

FIGS. 81 and 82 are cross-sectional plan views taken, respectively,along the lines 81-81 and 82-82 in FIG. 80.

FIG. 83 is a flow chart illustrating a method for operating the stainershown in FIGS. 79-82 in accordance with an embodiment of the presenttechnology.

FIGS. 84 and 85 are plots of average temperature and average airflowvelocity, respectively, within the internal environment relative to timeduring the method corresponding to the flow chart shown in FIG. 83.

FIG. 86 is a flow chart illustrating a portion of the methodcorresponding to the flow chart shown in FIG. 83 during which specimenson slides carried by a slide carrier are processed within the internalenvironment.

FIGS. 87 and 88 are plots of average temperature and average airflowvelocity, respectively, within the internal environment relative to timeduring the method corresponding to the flow chart shown in FIG. 86.

FIG. 89 is a perspective view of a liquid supply in accordance with oneembodiment of the present technology.

FIG. 90 is an isometric exploded view of a container in accordance withone embodiment of the present technology.

FIG. 91 is a partial cross-sectional side elevation view of thecontainer of FIG. 90.

FIG. 92 is an isometric view of a waste container in accordance with oneembodiment of the present technology.

FIG. 93 is a cross-sectional side elevation view of a sensor of thewaste container of FIG. 92.

DETAILED DESCRIPTION

Increasing the consistency and controllability of certain attributes(e.g., stain intensity) of histologically processed specimens is oftendesirable. Processing time (i.e., the duration of a given histologicalprocess) and processing temperature (i.e., the temperature at which agiven histological process is carried out) are two variables that affectmost, if not all, of these attributes. Automated histological systemsconfigured in accordance with at least some embodiments of the presenttechnology include features that facilitate consistency and/orcontrollability of processing time and/or processing temperature. Forexample, at least some of these systems include stainers havingprocessing heads capable of executing precisely controlled liquiddispensing and removing operations. These stainers can also haveinternal environments that can be maintained at elevated baselinetemperatures. The performance (e.g., with respect to quality and/orversatility) of these and other systems configured in accordance withembodiments of the present technology is expected to far exceed that ofconventional counterparts. Furthermore, systems configured in accordancewith at least some embodiments of the present technology can includefeatures that provide other desirable enhancements, such as reducedprocessing costs, reduced waste generation, and increased throughput.

Processing liquids selected or formulated in accordance with at leastsome embodiments of the present technology can differ from correspondingconventional processing liquids. For example, processing liquidsselected or formulated in accordance certain embodiments of the presenttechnology are less volatile than corresponding conventional liquids.For this reason and/or other reasons, these liquids may be well suitedfor use in stainers maintained at elevated baseline temperatures. Incontrast, corresponding conventional liquids may tend to evaporate atunacceptably high rates when used in these stainers. Evaporation ofprocessing liquids in automated histological systems is generallyundesirable. Furthermore, processing liquids selected or formulated inaccordance with embodiments of the present technology can be less toxicthan corresponding conventional processing liquids. This can facilitatedisposal of the processing liquids and/or reduce or eliminate therelease of noxious fumes from systems in which the processing liquidsare used. In at least some cases, some or all processing liquids usedwith an automated histological system configured in accordance with anembodiment of the present technology have relatively low concentrationsof monohydric alcohol (e.g., ethanol). For example, these processingliquids can include greater volumetric concentrations of polyol (e.g.,propylene glycol) than of monohydric alcohol. This can reduceevaporation, enhance certain aspects of specimen processing, anddecrease process complexity, among other advantages. Furthermore,processing liquids selected or formulated in accordance with embodimentsof the present technology can include other features that provide theseand/or other desirable enhancements.

Specific details of several embodiments of the present technology aredisclosed herein with reference to FIGS. 1-93. It should be noted thatother embodiments in addition to those disclosed herein are within thescope of the present technology. For example, embodiments of the presenttechnology can have different configurations, components, and/orprocedures than those shown or described herein. Moreover, a person ofordinary skill in the art will understand that embodiments of thepresent technology can have configurations, components, and/orprocedures in addition to those shown or described herein and that theseand other embodiments can be without several of the configurations,components, and/or procedures shown or described herein withoutdeviating from the present technology.

Selected Examples of System Architecture

FIG. 1 is an elevation view an automated slide processing system 2(“system 2”) in accordance with an embodiment of the present technology.The system 2 can include an access port 3 and an input device in theform of a touch screen 5. A user can load the system with slide-carryingtrays (“slide trays”), such as by placing the slide trays into theaccess port 3. A given slide tray can carry slides respectively carryingspecimens to be processed. Before, during, or after loading the system,the user use the touch screen 5 to select processes (e.g., protocols,recipes, etc.) to be performed on the specimens. The system 2 can thenautomatically process the specimens, apply coverslips to the slides, andreturn the slide tray to the access port 3. Thereafter, the coverslippedslides (e.g., slides carrying coverslips permanently coupled to theslides) can be retrieved from the access port 3 for subsequent analysis,pathologist interpretation, and/or archiving.

FIG. 2 is a side elevation view of system 2 showing some of its internalcomponents. The system 2 can include a housing 7 and modules (e.g.,workstations) 4, 6, 8 and 10 disposed within the housing 7. Also withinthe housing 7, the system 2 can include a transporter 12, a liquidsupply 14, a pressurization apparatus 16, and a controller 18. Thehousing 7 can maintain a generally contaminate-free internal environmentand/or help maintain a desired internal temperature suitable foroperating one or more of the modules 4, 6, 8, 10. A slide tray holdingspecimen-bearing slides can be carried by the transporter 12 between themodules 4, 6, 8, 10 to dry the specimens, stain the specimens, and applycoverslips to the slides. The specimens can be individually processed onslide without the use of shared baths of processing liquids. In thisway, cross-contamination, carryover of processing liquids, excessivewaste (e.g., liquid waste), inconsistent processing liquid performanceand other disadvantages of dip and dunk machines can be reduced oravoided. Furthermore, stain intensity and/or other post-processingattributes of the specimens can be highly controllable and preciselyexecutable. The transporter 12 and modules 4, 6, 8 and 10 can be underthe control of the controller 18, which can be controlled by a userusing the touch screen 5 (FIG. 1).

The module 4 can be a heater apparatus in the form of a dryer (“dryer4”), modules 6 can be stainers (“stainers 6”), module 8 can be acoverslipper (“coverslipper 8”), and module can be a heater apparatus inthe form of a curing unit (“curing unit 10”). The modules can bearranged in a vertical stack with the dryer 4 and curing unit 10positioned higher than the stainers 6. This can be useful, for example,because the dryer 4 and the curing unit 10 can generate heat, which canbe released through the top of the housing 7. The stainers 6 can beconnected to a fluidics manifold 19 that supplies liquids, such asstaining reagent (e.g., hematoxylin reagent) and counterstaining reagent(e.g., eosin reagent) from the liquid supply 14. The fluidics manifold19 can include, without limitation, one or more lines, valves, orifices,sensors, pumps, filters, and/or other components capable of controllablydelivering liquid. An electronics manifold (not shown) cancommunicatively couple the modules to the controller 18 to provide powerto and control over components of the modules and components thereof. Inone embodiment, individual modules are connected to the fluidicsmanifold 19 and the electrical manifold through common interfaces andplugs, respectively. The interchangeability afforded by using commoninterfaces and plugs may make it possible to add and remove modulesquickly and easily, thereby facilitating system reconfiguration,maintenance, and/or repair.

The transporter 12 can robotically move slide trays from module tomodule in an efficient manner so to enhance system throughput. Thetransporter 12 can comprise, without limitation, one or more elevators(e.g., rail and carriage assemblies), robotic arms, motors (e.g.,stepper motors, drive motors, etc.), tray interfaces or holders (e.g.,forks, clamps, etc.), and/or sensors, as well as other components forproviding motion. In at least some embodiments, the transporter 12includes an elevator and an inserter (e.g., an X-Y shuttle table) tofunction as an X-Y-Z transport mechanism (e.g., X-left to right; Y-frontto back; Z-up and down). Sensors (not shown) can be placed adjacent tothe transporter 12 to detect the position of the transporter 12 and usedto index the transporter 12 at sensing locations to provide preciseslide-tray positioning.

Sensors can be located at various locations throughout the system 2,including on the transporter 12, within the modules, and on the slidetrays. In some embodiments, sensors (including, without limitation,strain gauges, accelerometers, contact sensors, optical sensors, orother sensing devices capable of sensing certain events) can be used todetect collisions, impacts, or other events within the system 2. Thesensors can output one or more signals that are received by thecontroller 18, which can determine whether a given event requires usernotification or other action. For example, if an unexpected slide trayimpact is detected, the controller 18 can alert a user to open thehousing 7 to visually inspect the tray to determine whether slides areproperly positioned on the tray. Sensors can be mounted to a ceiling 13of the housing 7 to help prevent contact between the ceiling 13 and theslide trays and/or slides.

A holding station 23 with vertically spaced apart shelves 24 (oneidentified) can be positioned adjacent to and in front of thetransporter 12. An uppermost shelf 24 can be positioned underneath thedryer 4 and a lowermost shelf can be positioned above the access port 3.The transporter 12 can robotically move slide trays from the shelves 24to the dryer 4 to dry wet biological specimens, bake biologicalspecimens onto slides, or otherwise thermally process specimen-bearingslides. In some embodiments, the dryer 4 convectively heatsspecimen-bearing slides while holding the slides at orientations thatfacilitate drying. High convective flow rates can be used to providesubstantially uniform heating of the specimen-bearing slides to reduce(e.g., minimize) temperature differences across the specimens and/orslides due to, for example, the respective locations of the specimensand/or the slides in a slide tray.

The controller 18 can be part of a laboratory information managementsystem that can be connected, for example, to additional automatedstaining systems. The controller 18 can include, without limitation, oneor more printed circuit boards including any number of microprocessorsthat control, for example, the supply of processing liquids to themodules and module operation. Additionally or alternatively, printedcircuit boards, microprocessors, power sources, memory, readers (e.g.,label readers) and can be part of the individual modules and incommunication with the controller 18 or another controller, such as aremote controller. The controller 18 can command system components andcan generally include, without limitation, one or more centralprocessing units, processing devices, microprocessors, digital signalprocessors (DSPs), application-specific integrated circuits (ASICs),readers, and the like. To store information, the controller 18 caninclude, without limitation, one or more storage elements 21(illustrated in phantom), such as volatile memory, non-volatile memory,read-only memory (ROM), random access memory (RAM), or the like. Thestored information can include heating programs, staining programs,curing programs, coverslipping programs, optimization programs,specimen-processing programs (e.g., arbitrary user-defined sets ofoperations and/or pre-defined sets of operations), calibration programs,indexing programs, purge/prime programs, or other suitable executableprograms. Specimen-processing programs can include recipes or protocolsthat can be selected based on user preferences, such as pathologistpreferences. Optimization programs can be executed to optimizeperformance (e.g., enhance heating, reduce excess processing-liquidconsumption, increase productivity, enhance processing consistency, orthe like). System processing may be optimized by determining, forexample, an optimum schedule to (1) increase processing speeds, (2)reduce the time of heating cycles in the dryer 4 and/or in the curingunit 10, (3) increase throughput (e.g., increase the number of slidesprocessed in a certain length of time), (4) improve stain consistencyand/or quality, and/or (5) reduce liquid waste.

The liquid supply 14 can include slots for holding supply containers 27(one identified) and can include container identifiers, such asidentifiers with of RFID antennae that can read RFID tags associatedwith the supply containers 27. The supply containers 27 can include,without limitation, one or more human readable labels, machine readablelabels (e.g., a barcode to be read by the system 2), or other types oflabels. For example, the supply containers 27 can include RFID tagsencoded with information (e.g., container contents information,manufacture dates, expiration dates, etc.) about a particular processingliquid. One example of a container is discussed in connection with FIGS.90 and 91, and one example of a liquid supply is discussed in connectionwith FIG. 89. The liquid supply 14 can also include, without limitation,sensors (e.g., pressure sensors, temperature sensors, etc.), pumps(e.g., pneumatic pumps), valves, filters, lines, and/or other fluidiccomponents that can cooperate to supply liquids to the stainers 6, forexample.

The pressurization apparatus 16 can be located below the liquid supply14 and can include a plurality of pumps, compressors, vacuum devices(e.g., a blower), and/or other devices capable of pressurizing fluidsand/or providing a vacuum (including a partial vacuum). Pressurized aircan be delivered to, for example, air knives of the stainers 6, andvacuum level pressures can be used by liquid removal devices of thestainers 6.

Liquid waste can be delivered through lines and into waste containers32, 34. This waste can be generated within the system 2 from a varietyof sources. For example, liquid waste collected in the slide trays canbe removed and routed to the waste containers 32, 34. Periodicallyremoving this liquid waste can be useful to keep the waste from spillingout of the slide trays during handling. In the dryer 4, the slide traysmay collect mounting media (e.g., water), which can be sucked from theslide trays and pumped to one of the waste containers 32, 34. In thestainers 6, the slide trays can collect processing liquids that fall offthe slides, as well as liquids that inadvertently drip from nozzles ofdispenser apparatuses. In the coverslipper 8, the slide trays cancollect coverslipping liquids used to apply coverslips to the slides.The mounting media, processing liquids, coverslipping liquids, and anyother collected waste liquids can be pumped to the waste containers 32,34. A door 35 (FIG. 1) of the housing 7 can be opened to access andempty the waste containers 32, 34.

In operation, the slide trays can be loaded into the system 2 via theaccess port 3. Referring now to FIG. 2, the transporter 12 can retrievethe slide trays from the access port 3 and transport the slide trays todesired locations. The system 2 can individually process a particularspecimen-bearing slide and/or slide tray according to an arbitraryuser-defined set of operations, a pre-defined set of operations, orother sets of operations. The slide trays can be transported to aninterrogation station where the slides in the tray are analyzed bydetectors (e.g., optical sensors, cameras, etc.). The slide tray maythen then moved to the dryer 4 where specimens are dried and/or adheredto the slides. In some processes, the dryer 4 can help remove paraffinfrom paraffin-embedded specimens by melting and spreading the paraffinacross the surfaces of the slides. The resulting thin layers ofparaffin, having greater surface area once spread across the slides, maybe more easily removed by deparaffinizing liquid applied to the slideswithin the stainers 6. Once the specimens and/or slides have been atleast partially dried, the slide tray can be moved to one of thestainers 6 where the biological specimens are processed. The stainers 6can perform deparaffinizing, staining, conditioning (e.g., solventexchange), and other specimen processing operations by individuallyapplying fresh liquids to the specimens. This can facilitate controlover the post-processing characteristics of the specimens. The stainers6 can controllably dispense fresh processing liquids onto the slideswithout splattering onto adjacent slides and can controllably remove theprocessing liquids from the slides. The controlled dispensing/removalcan be used to efficaciously process specimens while also reducingvolumes of liquid waste by, for example, minimizing or otherwiselimiting volumes of liquid waste collected by slide trays. Theillustrated system 2 includes three stainers 6 that respectively provideparallel processing of three slide trays to increase system throughput,but a greater or lesser number of stainers can be used to prevent unduelimiting of the throughput of the system based on operation of thestainers 6.

As used herein, the terms “reagent” and “processing liquid” refer to anyliquid or liquid composition used in a specimen processing operationthat involves adding liquid or liquid composition to a slide. Examplesof reagents and processing liquids include solutions, emulsions,suspensions, and solvents (either pure or mixtures thereof). These andother examples can be aqueous or non-aqueous. Further examples includesolutions or suspensions of antibodies, solutions or suspensions ofnucleic acid probes, and solutions or suspensions of dye or stainmolecules (e.g., H&E staining solutions, Pap staining solutions, etc.).Still further examples include solvents and/or solutions fordeparaffinizing paraffin-embedded biological specimens, aqueousdetergent solutions, and hydrocarbons (e.g., alkanes, isoalkanes andaromatic compounds such as xylene). Still further examples includesolvents (and mixtures thereof) used to dehydrate or rehydratebiological specimens. The stainers 6 can receive a wide range ofreagents and processing liquids from the containers 27.

The term “staining” is used herein generally refers to any treatment ofa biological specimen that detects and/or differentiates the presence,location, and/or amount (such as concentration) of a particular molecule(such as a lipid, protein or nucleic acid) or particular structure (suchas a normal or malignant cell, cytosol, nucleus, Golgi apparatus, orcytoskeleton) in the biological specimen. For example, staining canprovide contrast between a particular molecule or a particular cellularstructure and surrounding portions of a biological specimen, and theintensity of the staining can provide a measure of the amount of aparticular molecule in the specimen. Staining can be used to aid in theviewing of molecules, cellular structures and organisms not only withbright-field microscopes, but also with other viewing tools, such asphase contrast microscopes, electron microscopes, and fluorescencemicroscopes. Some staining performed by the system 2 can be used tovisualize an outline of a cell. Other staining performed by the system 2may rely on certain cell components (such as molecules or structures)being stained without or with relatively little staining other cellcomponents. Examples of types of staining methods performed by thesystem 2 include, without limitation, histochemical methods,immunohistochemical methods, and other methods based on reactionsbetween molecules (including non-covalent binding interactions), such ashybridization reactions between nucleic acid molecules. Particularstaining methods include, but are not limited to, primary stainingmethods (e.g., H&E staining, Pap staining, etc.), enzyme-linkedimmunohistochemical methods, and in situ RNA and DNA hybridizationmethods, such as fluorescence in situ hybridization (FISH).

After processing the specimens, the transporter 12 can transport theslide trays from the stainer 6 to the coverslipper 8. The coverslipper 8can apply solvent to the slides and can place coverslips withpre-applied adhesive onto the slides. In some embodiments, the slidetray holds a plurality of slides in, for example, a substantiallyhorizontal position, and coverslips are individually added to theslides. In one embodiment, the coverslipper is substantially asdescribed in U.S. Patent Application Publication No. 2004/0092024A1 orU.S. Pat. No. 7,468,161, which are incorporated by reference herein intheir entireties. The coverslippers described in U.S. Patent ApplicationPublication No. 2004/0092024A1 or U.S. Pat. No. 7,468,161 and theiroperation can be implemented to enhance coverslip handling by, forexample, detecting broken coverslips, facilitating single coverslippickup, increasing coverslipper placement precision, and/or increasingsystem throughput.

Once coverslips are placed onto the slides, the transporter 12 cantransport the slide tray from the coverslipper 8 to the curing unit 10where coverslips are cured onto the slides (at least partially) and thetray itself is dried (at least partially) if the tray has collectedliquid. During curing, the slides can be held in substantiallyhorizontal positions to expose surface areas of the coverslips andslides to convective flows. This may facilitate quick and efficientcuring of adhesive. Even if coverslipping solvent underneath a givencoverslip is not completely removed, a skin of adhesive can form aroundthe coverslip that holds the coverslip in place during subsequenthandling by, for example, a health care professional, such as apathologist. In other embodiments, the curing unit 10 can include one ormore radiant heaters or conductive heaters, as well as combinations ofconvective heaters and radiant or conductive heaters. Once the slidesare coverslipped, the slide tray can be moved from the curing unit 10back to the access port 3 for retrieval.

The system 2 can have any number of modules arranged in any suitablerelationship relative to each other. In the illustrated embodiment,three stainers 6 and the curing unit 10 are positioned substantiallydirectly above and below one another in a vertical stack. Additionallyor alternatively, modules can be arranged side-by-side in a horizontalconfiguration (e.g., the dryer 4 positioned next to the curing unit 10).The modules can also be arranged in a sloped vertical stack withworkstations arranged side-by-side at any intermediate level in thesloped stack. Examples of modules that can be included in the disclosedautomated slide processing systems include, but are not limited to, aheater apparatuses (e.g., convection or radiant heaters), a reader(e.g., code reader), a stainer module, a coverslipper module, and acombination module, such as a combined dryer and deparaffinizer, acombined deparaffinizer/stainer, a combineddeparaffinizer/stainer/solvent exchanger), and other types ofworkstations (including workstations disclosed in U.S. Pat. No.7,468,161) that can perform one or more slide processing operations(such as two or more) in a single workstation. Example heater apparatusare discussed in connection with FIGS. 3-14 and example stainers arediscussed in connection with FIGS. 15-88. Additional modules can beadded to the automated slide processing system 2 to provide any numberof functionalities for automated processing of specimens with minimal orno human intervention during normal operation.

The slide trays may have any suitable shape, and the slides held in agiven slide tray can be arranged in any suitable manner to hold anysuitable number of slides, for example, 5 or more slides, 10 or moreslides, 20 or more slides, or 30 or more slides. Several examples ofslide trays of different shapes and holding capacities are disclosed inU.S. Pat. No. 7,468,161, which is incorporated by reference in itsentirety. In some embodiments, the slide trays are generally rectangulartrays configured to hold two rows of slides that are held side-by-sideon both sides of the central long axis of the slide tray so that thelong dimensions of the slides are disposed outward from the long centralaxis of the tray. The rectangular trays can have a bottom and sidewallsthat define a reservoir for liquid collection. In other embodiments, theslide tray is a circular slide tray configured to hold slides in radialpositions in which the long dimensions (or longitudinal axes) of theslides are disposed inward from the outer edge of the tray toward thecenter of the tray. In yet other embodiments, the tray can be agenerally square tray configured to hold slides in two or three rows.The configuration of the slide tray can be selected based on thedimensions of the slides, dimensions of the modules, and/or theconfiguration of the transporter 12.

The slide trays can hold specimen slides in a spaced apart arrangementand in substantially horizontal positions. Holding all the slides inseparation and in essentially the same plane (e.g., a horizontal planeduring staining) can limit or prevent cross-contamination of slidesduring, for example, drying, deparaffinizing, staining, washing andsolvent exchanging, and other acts that involve dispensing liquids ontoslide surfaces. Although the terms “slide tray” or “tray” are usedherein for ease of reference to items that carry slides, unless thecontext clearly indicates otherwise, other slide carriers capable ofholding an array of slides can be utilized. The system 2 can be usedwith a variety of slide carriers that have, without limitation, slideretainers (e.g., clamps, suction cups, etc.), slide standoffs, suctiondevices (e.g., tubes, nozzles, etc.) used to remove liquids from thetrays, or other features for holding, manipulating, or otherwiseprocessing slides.

The term “slide” refers to any substrate (e.g., substrates made, inwhole or in part, glass, quartz, plastic, silicon, etc.) of any suitabledimensions on which a biological specimen is placed for analysis, andmore particularly to a “microscope slide” such as a standard 3 inch by 1inch microscope slide or a standard 75 mm by 25 mm microscope slide.

Examples of biological specimens that can be placed on a slide include,without limitation, a cytological smear, a thin tissue section (such asfrom a biopsy), and an array of biological specimens, for example atissue array, a DNA array, an RNA array, a protein array, or anycombination thereof. Thus, in one embodiment, tissue sections, DNAsamples, RNA samples, and/or proteins are placed on a slide atparticular locations.

The term “biological specimen” refers to any specimen (e.g., sample)including biomolecules (e.g., proteins, peptides, nucleic acids, lipids,carbohydrates, and combinations thereof) that is obtained from (orincludes) any organism, including viruses. Biological specimens caninclude tissue samples (e.g., tissue sections), cell samples (e.g.,cytological smears such as Pap or blood smears or samples of cellsobtained by microdissection), samples of whole organisms (e.g., samplesof yeast, bacteria, etc.), or cell fractions, fragments or organelles,such as those obtained by lysing cells and separating their componentsby centrifugation or otherwise. Other examples of biological specimensinclude, without limitation, blood, serum, urine, semen, fecal matter,cerebrospinal fluid, interstitial fluid, mucous, tears, sweat, pus,biopsied tissue (e.g., obtained by a surgical biopsy or a needlebiopsy), nipple aspirates, milk, vaginal fluid, saliva, swabs (e.g.,buccal swabs), or any material containing biomolecules derivedtherefrom.

Selected Examples of Drying and Curing Ovens and Associated Methods

FIG. 3 is a cross-sectional perspective view of a heater apparatus inthe form of a dryer apparatus 1100 (“apparatus 1100”) in a closedconfiguration holding a slide carrier 1200 configured in accordance withan embodiment of the present technology. Generally, the apparatus 1100can heat a flow of gas that becomes a heated turbulent gas flow forpromoting a generally uniform heat distribution across the flow. Theturbulent gas flow can be converted to a laminar gas flow that flowsacross and heats specimen-bearing slides S (one identified) carried bythe slide carrier 1200. The specimen-bearing slides S can bevertically-oriented to promote draining of liquid, such as residualmounting media (e.g., water), from the slides S. The upwardly directedlaminar gas flow can flow across the specimens to inhibit, limit, orsubstantially prevent downward movement of the specimens relative to theslides S due to, for example, gravity while the specimens dry.

The apparatus 1100 can include a housing 1122, a blower 1110, and aheater 1116. The housing 1122 can have one or more walls 1119 and a doorassembly 1101 that define an interior space 1123. The interior space1123 can be a chamber divided by a septum 1112 into a back chamber 1142and a carrier-receiving or front chamber 1140 (“front chamber 1140”)that are fluidly connected to form a circulation loop 1121 within thehousing 1122. The cross-sectional area (i.e., the area generallyperpendicular to the direction of the gas flow) of the front chamber1140 can be less than the cross-sectional area of the back chamber 1142such that a relatively high speed flow travels over the slides S while arelatively low speed flows travels along the back chamber 1142. The doorassembly 1101 can move the slide carrier 1200 into a vertically-orientedposition within the front chamber 1140 to convectively heat thespecimen-bearing slides S. The blower 1110 can include, withoutlimitation, one or more fans, pumps, or other pressurization devicessuitable for forced flow convection. In some embodiments, the blower1110 is positioned along the circulation loop 1121 and is configured todirect the gas flow towards the heater 1116.

The heater 1116 can be configured to raise an average temperature of thegas flowing along the circulation loop 1121. As the gas flows along theheater 1116, the heater 1116 can transfer thermal energy to the gas flowand can be positioned within the back chamber 1142 opposite from anupper row of slides (separated by the septum 1112) to improve heating ofan upper row of slides S. Such positioning of the heater 1116 can offsetthe potential reduction in the temperature of the gas passing over theupper row of slides caused by evaporation of liquid on a lower row ofslides. In some embodiments, the heater 1116 can include, withoutlimitation, one or more resistive heater elements and one or more heattransfer elements (e.g., fins, tubes, etc.). In other embodiments, theheater 1116 can include both a resistive heater and a non-resistiveheaters, such as Peltier devices.

The apparatus 1100 can include flow modifiers configured to alter thecharacteristics of the gas flow along various portions of thecirculation loop 1121. For example, as shown in FIG. 3, the apparatus1100 can include a flow modifier in the form of a turbulence promoter1118 positioned downstream of the heater 1116. The turbulence promoter1118 can include one or more baffles, perforated plates, ribs, bumps,grooves and/or any structure configured to create eddies, swirling, orother generally turbulent or chaotic states of gas motion. As usedherein, “turbulent” refers to a gas flow having a Reynolds numbergreater than 4,000. By way of example, the majority of the gas flow in aturbulent flow portion 1143 along a substantial majority (e.g., at least90%, 95%, or 98%) of the cross-sectional area perpendicular to thedirection of flow can have a Reynolds number greater than 4,000. In someembodiments, the turbulence promoter 1118 extends between the septum1112 and the back wall 1119 and across the back chamber 1142. In otherembodiments, the turbulence promoter 1118 can be positioned along aninterior surface 1151 of the housing 1122 and/or a surface of the septum1112 and extend into, but not necessarily across, the circulation loop1121. The turbulent gas flow created by the turbulence promoter 1118 caninduce mixing of gas along the turbulent flow portion 1143 of the backchamber 1142, thereby improving heat transfer efficiency by, forexample, doubling or tripling the heat transfer efficiency within thecirculation loop 1121. In some embodiments, the turbulence promoter 1118is configured to produce sufficient turbulence that the gas flow exitingthe turbulent flow portion 1143 has a substantially uniform temperatureacross the flow (i.e., a substantially uniform temperature in adirection perpendicular to the direction of flow). In other embodiments,the flow modifier can have other configurations to promote, for example,mixing of the gas flow.

Additionally, the apparatus 1100 can include a flow modifier in the formof a laminar flow promoter 1114 positioned downstream of the turbulentflow portion 1143. The laminar flow promoter 1114 can include one ormore guide vanes, tapered channels, arcuate surfaces, and/or anystructure configured to create a substantially laminar gas flow. As usedherein, “laminar flow” or “substantially laminar flow” refers to a gasflow having a Reynolds number less than 2,100. The circulation loop 1121can have one or more laminar flow portions 1156. In some embodiments,the majority of the gas flow along a majority (e.g., at least 60%) ofthe cross-sectional area perpendicular to the direction of flow has aReynolds number less than 2,100. For example, the portion of thecirculation loop 1121 containing the laminar flow promoter 1114 (e.g.,between the turbulent flow portion 1143 and the front chamber 1140) canbe a laminar flow portion. Also, at least a portion of the front chamber1140 (e.g., between the specimen-bearing faces of the slides S and theseptum 1112) can be a laminar flow portion. In some embodiments, theapparatus 1100 can have a transitional gas flow (e.g., a gas flow havinga Reynolds number between 2,100 and 4,000) in at least a portion of theturbulent and/or laminar flow portions.

As shown in FIG. 3, the laminar flow promoter 1114 can be positioned ata bend 1153 in the circulation loop 1121 to guide the heated gas fromthe turbulent flow portion 1143 to the front chamber 1140. In someembodiments, the laminar flow promoter 1114 can be a plurality of spacedapart, arcuate members 1145 a, 1145 b, 1145 c that can reduce the headloss around the bend 1153. Once downstream of the arcuate members 1145a-1145 c, the gas can flow upwardly along the lengths of the slides S(e.g., substantially parallel to longitudinal axes A_(S) of the slides S(one identified)) to, for example, evaporate liquid on the slides S,thermally process the specimens (e.g., melt wax in the specimen), and/ordry the specimens (as discussed in greater detail below with referenceto FIGS. 6A-7). In other embodiments, the laminar flow promoter 1114 canbe positioned anywhere along the circulation loop 1121, such as along arelatively straight section of the circulation loop 1121.

In some embodiments, the laminar flow promoter 1114 can also acceleratethe gas flow to produce a relatively high speed laminar flow andincrease the rate of convective heating and/or evaporation rate. Forexample, in particular embodiments, the arcuate members 1145 a-1145 ccan define channels 1147 (one identified) that narrow in the downstreamdirection. As the gas flows through the channels 1147, the flow can beaccelerated to produce a high speed flow. In some embodiments, a ratioof the flow speed in the front chamber 1140 to the flow speed in theback chamber 1142 is equal to or greater than 2, 3, 4, 5, or 6. Theratio can be selected based on the desired specimen heating rates,evaporation rates, or the like.

One exemplary drying process is discussed below with reference to FIGS.4A-9. Generally, the slide carrier 1200 can be moved to a loadingposition while the slide carrier 1200 holds the slides S. The slidecarrier 1200 is robotically moved from the loading position to aprocessing position to move the slide carrier 1200 into the circulationloop 1121. The processing position can be angled relative to the loadingposition to facilitate drying of the specimens. The specimen-bearingmicroscope slides S are heated while the slide carrier 1200 is held atthe processing position. Details of the drying process are discussedbelow.

FIG. 4A is a side view of the apparatus 1100 in an open configurationbefore a slide carrier 1200 (shown schematically) has been placed on thedoor assembly 1101 by the transporter 12 (shown schematically), and FIG.4B is an enlarged top perspective view of the door assembly 1101.Referring to FIGS. 3-4B together, the door assembly 1101 can be disposedat a front portion 1103 (FIGS. 4A and 4B) of the apparatus 1100 and caninclude a door 1102, an actuation device 1108, and a kinematic mount1104. The door 1102 is moveable between a closed configuration (e.g.,FIG. 3) and an open configuration (e.g., FIGS. 4A-4B). The door 1102 canhave an interior surface 1130 that faces an interior portion of thehousing 1122 (within the circulation loop 1121) when the door 1102 is inthe closed configuration and an exterior surface 1132 (FIG. 4A) thatfaces outwardly when the door 1102 is in the closed configuration. Thedoor 1102 can be automatically moved between the closed and openconfigurations via the actuation device 1108. If the apparatus 1100shuts down (e.g., during a power outage), a user can manually open thedoor 1102 to retrieve any slide carrier in the apparatus 1100.

The actuation device 1108 can pivotally couple the door 1102 to thehousing 1122. In some embodiments, the actuation device 1108 includes amount 1111, a drive device 1113 (FIG. 4B), and a rotatable arm 1107(FIG. 4A). The mount 1111 is connected to the housing 1122 such thatdrive device 1113 is capable of rotating the arm 1107 (FIG. 4A) about apin 1109 of the mount 1111. The drive device 1113 can include, forexample, one or more drive motors, stepper motors, or other devicescapable of rotating the arm 1107. The configuration of the actuationdevice 1108 can be selected based on the desired motion of the door1102.

The kinematic mount 1104 can be coupled to the door 1102 and can includesupports 1106 (one identified) configured to hold and stabilize theslide carrier 1200 at a wide range of positions, including a horizontalposition and a vertically-oriented position (for example, as shown inFIG. 3). The kinematic mount 1104 can also include one or more kinematicmount sensors 1105 configured to detect the presence and/or position ofthe slide carrier 1200. In some embodiments, the sensor(s) 1105 candetect the presence and/or position of the slide carrier 1200 and alsohelp inhibit or limit movement of the slide carrier 1200. For example,the sensor(s) 1105 can be magnetic sensors that are capable of detectingthe presence/position of the slide carrier 1200 via a magnetic force.The magnetic force can help prevent sliding of the slide carrier 1200relative to the kinematic mount 1104. Other types of mounts can be usedto hold the slide carrier 1200, if needed or desired.

Referring now to FIG. 4A, when the door 1102 is in the openconfiguration, the door 1120 can be substantially horizontal andconfigured to receive the slide carrier 1200 from the transporter 12.The term “substantially horizontal” with reference to the door assembly1101 generally refers to an angle within about +/−2 degrees ofhorizontal, for example, within about +/−1 degree of horizontal such aswithin about +/−0.8 degrees of horizontal. When the door 1102 issubstantially horizontal it can have an orientation such that theinterior 1130 and exterior 1132 surfaces of the door 1102 are generallyfacing up and down, respectively.

Once the transporter 12 delivers the slide carrier 1200 to the apparatus1100 vicinity, the transporter 12 can place the slide carrier 1200 ontothe kinematic mount 1104. At this point, both the transporter 12 and thekinematic mount 1104 can be engaged with the slide carrier 1200. Ifneeded, the transporter 12 can reposition the slide carrier 1200relative to the door 1102 and/or kinematic mount 1104 based on signalsreceived from the kinematic mount sensors 1105 and/or transportersensors (not shown). Once a desired positioning is achieved, thetransporter 12 relinquishes the slide carrier 1200 to the door assembly1101, as shown in FIG. 5.

Referring now to FIG. 5, the door 1102 in the open configuration cansupport the slide carrier 1200 in a substantially horizontal positionsuch that such that the largest surfaces of the slides (collectivelyreferred to as “S”) are generally facing up and down. In the illustratedembodiment, the slide carrier 1200 is shown including a first row 1201of slides S (one identified) and a second row 1203 of slides S (oneidentified). In other embodiments, however, the slide carrier 1200 cancontain more or less than two rows (e.g., a single row, three rows,etc.) and/or each row can include any number of slides (e.g., one, five,ten, twelve, etc.).

FIG. 6A is a cross-sectional side view of the apparatus 1100 after thedoor 1102 carrying the slide carrier 1200 has rotated upwardly to anvertically-oriented, closed configuration. FIG. 6B is an enlargedcross-sectional side view of a portion of the door assembly 1101 holdingthe slide carrier 1200. Referring to FIGS. 6A-6B together, the slidecarrier 1200 is enclosed within the housing 1122 and holds the slides Swithin the front chamber 1140 of the circulation loop 1121. The blower1110 pushes a gas (e.g., air or other suitable gas) over the heater1116, through and/or over the turbulence promoter 1118, through and/orover the laminar flow promoter 1114, and upwardly along thespecimen-carrying faces of the slides S to convectively heat extraneousliquid on the slides S and/or specimens carried by the slides S. Oncethe gas leaves the front chamber 1140, the gas can be re-circulated bythe blower 1110. In the illustrated embodiment, the gas flow movesthrough the circulation loop 1121 in a generally counterclockwisedirection. However, in other embodiments, the gas flow can be in aclockwise direction. The flow rate across the slides S can be generallyuniform, and can be between 1.8 m/s and 2.9 m/s (e.g., 2.8 m/s). Becausethe laminar gas flow can travel across the specimens without pushing thespecimens off the slides S, relatively high flow rates can be used. Ifthe flow rate is too low, extraneous liquid can remain on the slides,thereby allowing specimen migration (e.g., migration of a distance equalto or greater than 2 mm) and possibly staining. If the flow rate is toohigh, the gas flow can cause specimen migration (e.g., the gas can pushspecimens up the slides a distance equal to or greater than 2 mm) or insome instances, damage the specimens. The blower 1110 can selectivelyincrease or decrease the flow rate to achieve target processing (e.g.,evaporation rates, draining rates, etc.) while limiting or preventingspecimen migration and/or damage.

As discussed, drying of the specimens and/or slides is achieved byconvective heating using the heater 1116 and the blower 1110. Generally,the temperature of the gas flow within the circulation loop 1121 can bemaintained within a desired processing temperature range, such as arange of about 65° C. to about 80° C. (e.g., about 72-73° C.). As such,during the drying process the slides S and/or specimens are uniformlyheated such that at any point during the drying process, the temperatureof the individual slides S are within 5° C. of one another (includingnone, all or a subset of the slides being at substantially the sametemperature). Achieving an appropriate temperature can be advantageousbecause, for example, if the temperature is not low enough, the slidesand/or specimens may not be dried within the allotted time for thedrying process. Moreover, delivering a heated gas flow having an averagetemperature greater than 65° C. allows liquid within and/or underneathany wax or other material associated with the specimen to evaporate.

Referring now to FIG. 6B, the slide carrier 1200 can bevertically-oriented such that an axis A of the slide carrier 1200 and/orlongitudinal axes A_(S) (one identified) of the respective slides S areoriented at angle θ with respect to a horizontal plane H. As usedherein, “vertically-oriented” can refer to both an inclined/angledposition and a substantially vertical position. As used herein, an“inclined” or “angled” position refers to an orientation of the slidecarrier 1200 and/or slides S where the slide carrier 1200 and/orlongitudinal axis A_(S) (one identified) of the slides S are positionedat an angle θ that is between 70 degrees and 90 degrees (e.g., between77 degree and 84 degrees, 80 degrees, 90 degrees, etc.). As used herein,the term “substantially vertical” refers to an orientation of the slidecarrier 1200 and/or slides S where the slide carrier 1200 and/orlongitudinal axes A_(S) of the slides S are positioned at an angle θthat is within about +/−2 degrees of 90 degrees (including 90 degrees),for example, within about +/−1 degree of 90 degrees such as within about+/−0.8 degrees of 90 degrees. In either position, the first row 1201 ofslides is positioned vertically above the second row 1203 of slides suchthat a first end (1201 a, 1205 a) of each slide S is vertically above asecond end (1201 b, 1205 b) of the same slide S. The vertically-orientedslide carrier 1200 and/or slides S leverage the effect of gravity topull extraneous liquid off the slides S, thereby expediting the dryingtime.

Accordingly, the methods of the present technology are faster and moreeffective than conventional horizontal slide drying methods. Forexample, the drying time (i.e., the time between when the door 1102receives a slide carrier 1200 to when the transporter 12 removes theslide carrier 1200) can be between 2 minutes and 8 minutes (e.g., 3minutes, 4 minutes, 4.5 minutes, 5 minutes, etc.). For example, in oneembodiment the drying time can be 4 minutes and 52 seconds.

As discussed above, placing the slide carrier 1200 and/or slides S at avertically-oriented position during drying utilizes gravity toeffectively drain freestanding liquid on the mounting surfaces of theslides S. However, such a position also raises the possibility of aportion of a specimen in the first or upper row 1201 falling andcontaminating a slide S in the second or lower row 1203. Suchcross-contamination can impair subsequent analysis of the specimens.Accordingly, the position and the configuration of the slide carrier1200 can be adjusted to increase drying efficiency while avoiding orlimiting cross-contamination of the slides S. For example, FIG. 6B showsthe slide carrier 1200 and slides S in an inclined position. The labeledends of the slides S can be lower than their non-labeled ends such thatlabels (e.g., adhesive bar code labels) can inhibit or limit migrationof the specimens, if the specimens slide along the mounting surfaces ofthe slides. Thus, the labels can serve as physical barriers to keep thespecimens on the slides. In the illustrated embodiment, the slidecarrier 1200 includes one or more standoffs 1202 that separate theslides S from a surface 1204 of the slide carrier 1200 and the upper andlower slides S are horizontally spaced apart from one another. As such,liquid and/or specimens dripping from the upper row 1201 (depictedschematically as “D”) can fall directly downward onto the inclinedsurface 1204 of the slide carrier 1200, thereby avoidingcross-contamination of the lower slide S. In comparison, FIG. 7 showsthe slide carrier 1200 and slides S in a substantially verticalposition. Here, the slide carrier 1200 includes one or more barriers1602 between adjacent rows of slides S. As gravity pulls liquid off ofthe specimens and/or slides S, the liquid D can be caught by thebarrier(s) 1602, thereby preventing cross-contamination of the lowerslides S. The apparatus 1100 of FIGS. 3-6 can be modified to hold theslide carrier 1200 in such vertical orientation shown in FIG. 7.

Referring again to FIGS. 6A and 6B, ambient air can enter thecirculation loop 1121 via an opening 1606 to compensate for increasedhumidity within the housing 1122 due to evaporation of liquid of the wetspecimen-bearing slides S. The ambient air can have a relatively lowhumidity to help limit the humidity levels within the housing 1122 andthereby limit the humidity of the gas flow along the circulation loop1121. In some embodiments, the housing 1122 and/or sidewalls 1119 can besubstantially sealed to retain heat, although during the opening andclosing of the door 1102, gas and thermal energy is necessarilyexchanged with the external environment. This exchange allows therelative humidity within the interior space 1123 (FIG. 6A) and/orcirculation loop 1121 to equilibrate to an appropriate level andprevents moisture build-up as wet specimens are introduced.

Once the drying cycle is complete, the slide carrier 1200 is rotateddownwardly to a substantially horizontal position, as shown in FIG. 8.The transporter 12 can position itself adjacent to the door 1102 andsubsequently removes the slide carrier 1200 from the door assembly 1101.In some embodiments, the transporter 12 can have one or more extensionsthat project into the space between the slide carrier 1200 and theinterior surface 1130 of the door 1102 and engage a downward-facingsurface of the slide carrier 1200. At this stage, both the transporter12 and the kinematic mount 1104 can confirm engagement with the slidecarrier 1200. The transporter 12 can then automatically remove the slidecarrier 1200 from the kinematic mount 1104 and remove the slide carrier1200 from the immediate vicinity of the apparatus 1100. Feedback fromthe kinematic mount sensors 1105 and/or transporter sensors (not shown)can help guide the slide carrier removal process.

FIG. 9 is a cross-sectional side view of the apparatus 1100 in theclosed configuration, either after the slide carrier 1200 has beenremoved and the door 1102 has closed, or before the door 1102 opens toreceive a slide carrier 1200 from the transporter 12. Regardless, whenthe apparatus 1100 is in the closed configuration and the slide carrier1200 is not present, the heater 1116 can continuously or periodicallygenerate heat to maintain a desired standby temperature. Accordingly,when a subsequent slide carrier is introduced, there is less lag timefor the apparatus 1100 to recover to a desired operating temperature.

In some embodiments, the apparatus 1100 can include additional features.For example, in some embodiments, the apparatus 1100 can include heatersafety features. For example, the apparatus 1100 can include a heatsensor (not shown) on the heater 1116 that monitors a temperature of theheater 1116 and cuts power to the heater 1116 if the heater 1116 goesabove a specified temperature. Additionally, the heater 1116 itself caninclude a switch (e.g., a mechanical switch, electromechanical switch,etc.) that breaks the power circuit path if the heater 1116 goes above aspecified temperature. If the heater temperature returns to anappropriate level (e.g., below the specified temperature), the switchcan close the circuit, thereby enabling power delivery to the heater1116. The apparatus 1100 can include additional features to ensurerobust drying. For example, the apparatus 1100 can include one or morelayers of insulation that surround the housing 1122 and/or walls 1119 toretain heat and maintain proper heat distribution. Additionally, theapparatus 1100 can include one or more dehumidifying elements that limitthe humidity in the housing 1122 to enhance drying.

FIG. 10 is a perspective view of another embodiment of a heaterapparatus in the form of curing oven 1800 (“oven 1800”), in a closedconfiguration in accordance with an embodiment of the presenttechnology. The oven 1800 is generally identical to the apparatus 1100discussed in connection with FIGS. 3-9, except as detailed below. Theoven 1800 is configured to thermally process slides carrying coverslipsto cure the coverslips onto the slides in order to protect thespecimens. The oven 1800 can also alleviate any “carrier messiness”(i.e., free-standing extraneous liquid on the slides and/or slidecarrier) by heating the slides and/or slide carrier and evaporatingsuperfluous liquids (if present). Moreover, a substantially horizontalposition can be advantageous to help maintain the positioning orplacement of the coverslip on the slide (and likewise avoid coverslipmigration). The oven 1800 can include a housing 1822 having one or morewalls 1819 (FIG. 13) and a door assembly 1801. The door assembly 1801can hold a slide carrier to keep the coverslipped slides insubstantially horizontally orientations or other suitable orientations.An actuation device 1808 of the door assembly 1801 can include one ormore rails, carriages, drive mechanisms, or other components suitablefor vertically moving a door 1802 between a closed configuration (e.g.,FIG. 10) and an open configuration (e.g., FIG. 11).

One exemplary curing process is discussed below with reference to FIGS.11-14. Generally, a slide carrier 1200 is moved to the door assembly1801 while the slide carrier 1200 holds the coverslipped slides CS. Theslide carrier 1200 is moved robotically by the door assembly 1801 from afirst position (e.g., a horizontal lowered position) to a secondposition (e.g., a horizontal raised position) to move the slide carrier1200 into a circulation loop. The coverslipped slides CS are heatedwhile the slide carrier 1200 into a circulation loop. Details of theoven 1800 and the curing process are discussed below.

FIG. 11 is a perspective view of the curing oven 1800 in an openconfiguration before a slide carrier 1200 (shown schematically) has beenplaced on the door assembly 1801 by the transporter 12 (shownschematically). As shown in FIG. 11, the door assembly 1801 can bedisposed at a bottom portion 1803 of the oven 1800 and can include thedoor 1802 and the actuation device 1808. The door 1802 can have aninterior surface 1830 that faces an interior portion of the housing 1822and an exterior surface 1832 that faces outwardly. In some embodiments,including the illustrated embodiment, a kinematic mount 1804 is carriedby door 1802 and can include vertically oriented supports 1805 (oneidentified) configured to hold and stabilize a slide carrier 1200.

When the door 1802 is in the open configuration, the door 1802 can besubstantially horizontal and configured to receive the slide carrier1200 from the transporter 12. Once the transporter 12 delivers the slidecarrier 1200 to the curing oven 1800 vicinity, the transporter 12 placesthe slide carrier 1200 on the kinematic mount 1804. At this point, boththe transporter 12 and the kinematic mount 1804 can be engaged with theslide carrier 1200. Once a desired positioning is achieved, thetransporter 12 relinquishes the slide carrier 1200 to the door 1802, asshown in FIG. 12.

FIG. 13 is a cross-sectional side view of the curing oven 1800 after thedoor 1802 carrying the slide carrier 1200 has moved to the closedconfiguration. The slide carrier 1200 is enclosed within the housing1822 such that the coverslips and slides (together referred to as“coverslipped slides CS”) are exposed to laminar flow in the circulationloop 1821. In operation, the blower 1810 pushes a gas over the heater1816, through and/or over the vertically oriented turbulence promoter1818, through and/or over the laminar flow promoter 1814, and along thespecimen-carrying face of the coverslipped slides CS to convectivelyheat and/or cure the coverslipped slides CS. The flow rate across thecoverslipped slides CS can be generally uniform, and on average can bebetween 5 m/s and 7 m/s (e.g., 6 m/s). If the flow rate is too low, theflow rate may not efficiently cure the coverslips (i.e., cure theadhesive/glue carried by the coverslips) in the allotted processing timeand/or extraneous liquid may be left on the slide carrier 1200 and/orcoverslipped slides CS. Insufficient curing and/or drying can affectarchiveability (i.e., specimens can be stored upright in common slidedrawers without being stuck together, the stain can be retained and thecoverslip adhered to the specimen for at least 10 years). If the flowrate is too high, the flow rate can cause specimen or coverslipmigration or in some instances, damage the specimens. Accordingly, theflow rate can be selected based on the desired curing time whilelimiting or preventing migration of the specimens and/or coverslips.

Achieving an appropriate curing temperature can be advantageous because,for example, if the temperature rises above a specified threshold, thetemperature can affect the material properties of the coverslipmaterial. For example, without being bound by theory, it is believedthat going above certain temperatures can cause the coverslip to embeddeeply into the specimen, causing the coverslip to remain in thespecimen during de-stain and therefore negatively impact re-stains.Additionally, the higher the temperature in the oven 1800, the higherthe temperature of the slide carrier 1200, possibly requiring a “cooldown” period (or a longer cool down period) due to the fact that theslide carrier 1200 must be at an acceptable handling temperature whenexiting the oven 1800. A long cool down time can impact throughput.Also, maintaining an average curing temperature of less than 100° C. canbe advantageous to avoid burning or permanently damaging the specimensand/or slides. If the temperature is not low enough, the slides and/orspecimens may not be dried within the allotted time for the curingprocess. During the curing process the slide carrier 1200 can beenclosed or positioned within the circulation loop 1821 such that thecover-slipped slides CS are convectively heated. Accordingly, themethods of the present technology may be faster and more effective thanconventional horizontal drying methods. For example, the curing time(i.e., the time between when the door 1802 receives a slide carrier 1200to when the transporter 12 removes the slide carrier 1200) can bebetween 2 minutes and 8 minutes (e.g., 3 minutes, 4 minutes, 4.5minutes, 5 minutes, etc.). For example, in one embodiment the curingtime can be 4 minutes and 52 seconds. Generally, the average temperatureof the gas flow within the circulation loop 1821 can be between 90° C.and 110° C. However, other temperatures can be achieved to cure othertypes of adhesives used with coverslips.

Once the curing cycle is complete, the slide carrier 1200 is lowered forremoval by the transporter 12, as shown in FIG. 14. The transporter 12positions itself adjacent to the door 1802 and subsequently removes theslide carrier 1200 from the door assembly 1801.

In some embodiments, the transporter 12 can have one or more extensionsthat project into the space between the slide carrier 1200 and theinterior surface 1830 of the door 1802 and engage a downward-facingsurface of the slide carrier 1200. At this stage, both the transporter12 and the kinematic mount 1804 can confirm engagement with the slidecarrier 1200. The transporter 12 can then automatically remove the slidecarrier 1200 from the door assembly 1801 and transport the slide carrier1200 away from the immediate vicinity of the oven 1800. Feedback fromthe kinematic mount sensors and/or transporter sensors (not shown) canhelp guide the slide carrier removal process.

The curing oven 1800 can include additional features to ensure robustcuring. For example, the oven 1800 can include a layer of insulationthat surrounds the housing 1822 and/or sidewalls 1819 to retain heat andmaintain proper heat distribution. The housing 1822 and/or sidewalls1819 are substantially sealed to retain heat, although during theopening and closing of the door 1802, gas is necessarily exchanged withthe external environment. This exchange allows the relative humiditywithin the interior space 1823 and/or circulation loop 1821 toequilibrate to an appropriate level and prevents moisture build-up aswet specimens are introduced.

Selected Examples of Tray and Slide Handling in Stainers

FIG. 15 is an isometric view of a stainer module 2010 in an openconfiguration in accordance with an embodiment of the presenttechnology. The stainer module 2010 can include a tray handler 2020, ahousing 2022, and a dispenser apparatus 2024. The tray handler 2020 canmove a slide carrier in the form of a portable tray (not shown in FIG.15) through an opening 2023 of the housing 2022 and can position thetray underneath the dispenser apparatus 2024. The dispenser apparatus2024 can include four head or manifold assemblies 2018 a, 2018 b, 2018c, 2018 d (collectively “head assemblies 2018”) that providevalve-controlled, pressurized liquid delivery onto specimen-bearingmicroscope slides carried by the tray. To maintain a high processingthroughput, the head assemblies 2018 can be purged/primed while the trayremains positioned in the stainer module 2010. In dispense processes,the head assemblies 2018 can individually dispense predetermined volumesof fresh liquid onto slides and can remove the liquid from the slides toperform multi-step staining protocols. After processing the slides, thetray handler 2020 can move the tray out of the housing 2022.

The tray handler 2020 can include a tray holder transport mechanism 2030(“transport mechanism 2030”) and a tray holder in the form of akinematic mount 2040. The transport mechanism 2030 can include, withoutlimitation, a home flag and a relative encoder used to accuratelyposition the kinematic mount 2040. The kinematic mount 2040 can includearms 2041 a, 2041 b, 2041 c (collectively “arms 2041”), supports 2042 a,2042 b, 2042 c (collectively “supports 2042”), and a sensor 2046. Insome embodiments, the supports 2042 are mount balls connected to freeends of the arms 2041 to provide multi-dimensional constraints (e.g.,three-dimensional constraints). When the supports 2042 interface withthe tray, the sensor 2046 can detect the presence and/or position of thetray.

The transport mechanism 2030 and kinematic mount 2040 can minimize orlimit unintended motion of the tray that affects spacing between uppersurfaces of the slides and the head assemblies 2018. Increased spacingcan lead to splattering of liquids, whereas decreased spacing may resultin physical contact between the head assemblies 2018 and thespecimen-bearing slides. Splattering can lead to increased overallprocessing-liquid waste and understaining of specimens. If thesplattered liquid lands on adjacent slides, the specimens on theadjacent slides may be improperly stained. If the tray experiencessignificant pitch motion (e.g., pitch motion about the illustratedX-axis) and/or roll motion (e.g., roll motion about the illustratedY-axis), the head assemblies 2018 may contact and break slides and/ormay dislodge specimens. Unintended yaw motion (e.g., rotation about theillustrated Z-axis) of the tray can affect distances (e.g., X-axisdistances and Y-axis distances) between the edges of slides and the headassemblies 2018, which can result in processing liquid being directlydispensed into the tray. Because the desired volume of processing liquidis not delivered onto the slides, the specimens could be understained.The transport mechanism 2030 and kinematic mount 2040 can cooperate toinhibit, limit, or substantially eliminate unintentional motion of thetray (e.g., pitch motion, roll motion, and/or yaw motion) to inhibit,limit, or prevent one or more of the following: splattering of liquids,physically contact between the head assemblies 2018 and thespecimen-bearing slides, dislodging of specimens, and misaligningslides. By dispensing all (or substantially all) of the liquid directlyonto the slides, the liquids can be efficiently used, and the trays canremain substantially free of liquid throughout processing. As such,volumes of processing liquid used by the stainer module 2010 can besignificantly less than volumes of liquid used by conventional automatedslide stainers.

FIG. 16 is an isometric view of the stainer module 2010 after a traytransporter 2052 (shown schematically in phantom line) has placed a tray2050 onto the kinematic mount 2040. The tray transporter 2052 canreposition the tray 2050 based on signals from the sensor 2046 (FIG.15). The tray 2050 can hold slides in substantially horizontalorientations such that large surfaces of the slides are generally facingup and down. The term “substantially horizontal” generally refers to anangle within about +/−3 degrees of horizontal, for example, within about+/−1 degree of horizontal, such as within about +/−0.8 degrees ofhorizontal. Substantially horizontal also refers to ranges of smallangles from horizontal, for example, angles between about 0.1 degreesand 1.8 degrees from horizontal, such as angles between about 0.2degrees and about 1.2 degrees, for example angles between about 0.3degrees and about 0.8 degrees. In particular embodiments, an angle withupper surfaces of substantially horizontal slides relative to animaginary horizontal plane can be between about 0 degrees and about 3degrees along its short axis, and an angle with respect to the imaginaryhorizontal plane of between about 0 degrees and 2 degrees along its longaxis, again with the large surfaces of the slides generally facing upand down. The illustrated tray 2050 is capable of holding twenty slides,but it is shown holding only two slides 2053, 2054.

FIG. 17 is a bottom view of the stainer module 2010 holding the tray2050. The tray 2050 can include receiving features 2092 a, 2092 b, 2092c (collectively “receiving features 2092”) that interface withrespective supports 2042 a, 2042 h, 2042 c (FIG. 15). The receivingfeatures 2092 can be curved features, recesses, elongate slots, or otherfeatures that engage the supports 2042. In one embodiment, the receivingfeatures 2092 are partially spherical surfaces or arcuate grooves alongwhich the supports 2042 can slide to provide self-leveling of the tray2050, thereby keeping the tray 2050 substantially horizontal throughoutprocess.

The transport mechanism 2030 can include, without limitation, one ormore motors 2088 (e.g., drive motors, stepper motors, etc.) and a drivedevice 2089. The drive device 2089 can include, without limitation,rails, carriages, extendable arms, belts, chains, gear mechanisms, orcombinations thereof to provide translation of the tray 2050 along asingle axis or multiple axes. The transport mechanism 2030 can move thetray 2050 from a tray load/unload position (shown in FIGS. 16 and 17) toa processing position (shown in FIG. 18) with a chamber 2080 (FIG. 16)of the stainer module 2010. Due to small gaps between slide surfaces andthe head assemblies 2018, interferences are possible if a slide ismisaligned within the tray 2050 or if the tray 2050 is misaligned on thekinematic mount 2040, and such interferences may result in a shut-downevent. In the event of shut down of the stainer module 2010, a user canmanually manipulate the transport mechanism 2030 to position the tray2050 at an accessible position suitable for manually retrieving and/orrepositioning the tray 2050.

FIG. 18 shows the stainer module 2010 after the transport mechanism 2030has positioned the tray 2050 generally underneath the dispenserapparatus 2024. FIG. 19 is an isometric view of the dispenser apparatus2024 ready to process slides. The dispenser apparatus 2024 and the tray2050 can be moved in orthogonal directions to accurately position theslides relative to paths of travel of the head assemblies 2018. Afterprocessing the slides, the dispenser apparatus 2024 can be heldstationary or moved while the tray 2050 is indexed relative to the headassemblies 2018. The next four slides can be processed. This process canbe repeated until all the slides carried by the tray 2050 are processed.

Referring to FIGS. 19 and 20, a dispenser drive mechanism 2128 (“drivemechanism 2128”) can move the dispenser apparatus 2024 in the Y-axisdirection (i.e., a direction parallel to the illustrated Y-axis). Thepaths of travel of the head assemblies 2018 can be aligned with the longaxes of the slides extending in the Y-axis direction such that the headassemblies 2018 sweep along the lengths of the slides. In variousembodiments, the drive mechanism 2128 can include, without limitation,one or more rails, carriages, extendable arms, gear mechanisms, orcombinations thereof to provide translation along a single axis. In someembodiments, including the illustrated embodiment, the drive mechanism2128 includes a motor 2131 and a translator device 2132. The translatordevice 2132 includes a rail 2135 and a carriage 2136 (FIG. 19) movablealong the rail 2135. A frame 2108 of the dispenser apparatus 2024 cancarry the head assemblies 2018 and is coupled to the carriage 2136.

The head assemblies 2018 b, 2018 c can apply liquids to slidespositioned under an opening 2120 in a plate 2124, and the headassemblies 2018 d, 2018 a can apply liquids to slides positioned underan opening 2122 (FIG. 20) in the plate 2124. The transport mechanism2030 can move the tray 2050 in the X-axis direction (i.e., a directionparallel to the illustrated X-axis as indicated by arrows 2123, 2125 inFIG. 20) to sequentially position slides under the head assemblies 2018.Single-axis motion of the tray 2050 can facilitate lateral alignment ofthe slides with the head assemblies 2018. FIG. 21 shows the headassemblies 2018 c, 2018 d positioned above the tray 2050. FIG. 22A is adetailed view of the head assembly 2018 positioned above a slide 2160 ata processing zone 2170. (Hoses and other components of the stainermodule are not shown to avoid obscuring features in the images.) Thehead assembly 2018 can include a dispenser head 2141, valves 2143, 2145,and lines 2147. FIG. 22B shows tray 2050 moved to position another slideat the processing zone 2170.

FIGS. 23-26 are views of stages of applying substances to microscopeslides. Generally, slides can be sequentially positioned underneath andindividually processed by the head assemblies 2018. Slide processing isdiscussed in connection with a single head assembly 2018. However,multiple head assemblies 2018 can simultaneously or sequentially processslides in a similar manner. FIG. 23 shows twenty slides spaced apartfrom one another in two rows. When the tray 2050 is in substantiallyhorizontal orientation, the mounting areas of the slides can faceupwardly. However, the slides can be held in other arrangements and atdifferent orientations, if needed or desired.

Referring to FIGS. 22A and 24, the head assembly 2018 is ready toprocess slides at the processing zone 2170 (illustrated in phantomline). Each distributed dispense can form a relatively thick film (orpuddle) over any specimens on the slide 2160 (FIG. 22A) to incubate in adesired mode, such as a quasi-static mode. Each dispense, for example,can form a puddle having a shape at least partially maintained bysurface tension. In some embodiments, the head assembly 2018 can movelengthwise along the stationary slide 2160 at a speed in a range ofabout 1 inch/second to about 15 inches/second and can be accelerated upto 100 inch/second². Other speeds can be used to match liquid flow/valvetimes for dispensing or homing operations. For example, the headassembly 2018 can be moved relatively slowly (e.g., 1 inch/second toabout 2 inches/second) during homing operations, such as movement of thehead assembly 2018 to a home position. In other embodiments, the headassembly 2018 can move lengthwise along the slide 2160 while the slide2160 moves in the X-direction, Y-direction, and/or Z-direction. Forexample, the slide 2160 can be moved in the X-direction to periodicallyor continuously laterally reposition the slide 2160 during thedispensing process.

FIG. 25 shows the head assembly 2018 after processing the slide 2160.The head assembly 2018 can then process a slide 2210 position at theprocessing zone 2170, and after processing the slide 2210, the tray 2050can be moved in the X-direction (indicated by arrow 2192) to move theslides 2270, 2271 to the processing zone 2170. In some embodiments, traymovement can occur after the Y-axis motion of the head assembly 2018 hasbeen initiated or completed to minimize the potential of interferenceimpacts and/or to enhance system throughput. The head assembly 2018 canbe moved to a “safe” position to provide interference points that arespaced apart from the path of the tray 2050, which can be moved at aspeed selected to keep processing times as low as possible withoutcompromising controlled liquid distribution. For example, the tray 2050can move at a speed in a range of about 5 inches/second to about 6inches/second. Other speeds can be used, if needed or desired. FIGS. 22Band 26 shows one of the head assemblies 2018 ready to process the slide2270. A puddle 2240 is shown dispensed on a surface of the slide 2160.Each of the head assemblies 2018 can sequentially process the slideswithin a given quadrant. In some embodiments, the tray 2050 is movedduring the dispense process. For example, the trays 2050 can be movedrelative to the head assembly 2018 while the head assembly 2018dispenses liquid to form, for example, zig-zag shaped puddles (as viewedfrom above), serpentine puddles, or other shaped puddles. The movementof the tray 2050 and head assembly 2018 can be coordinated to produce awide range of different shaped puddles.

FIG. 27 is a view of the stainer module 2010 taken along line 27-27 ofFIG. 21 with tray not shown. FIG. 28 is a cross-sectional view of aliquid collector 2300 taken along line 28-28 of FIG. 27 and a front viewof two head assemblies 2018 d, 2018 c. Referring now to FIG. 27, theliquid collector 2300 can be a tray or a pan with spaced apartreservoirs 2310 a, 2310 b, 2310 c, 2310 d (collectively “reservoirs2310”) positioned to collect liquid from the head assemblies 2018 a,2018 b, 2018 c, 2018 d, respectively. The description of one reservoir2310 applies equally to the other reservoirs 2310, unless indicatedotherwise.

Referring now to FIG. 28, the reservoir 2310 d can include a drain 2314and a sloped surface 2330 for directing liquid to the drain 2314. Thedrain 2314 can be fluidically coupled to a waste module (or wastecontainer) or other component by one or more lines. Liquid can becontinuously or periodically drained from the reservoir 2310 d. In someembodiments, the reservoir 2310 d has a conical shape. In otherembodiments, the reservoir 2310 d has a frusto-conical shape, but thereservoir 2310 d can have other configurations. The head assemblies 2018can dispense liquid directly into the reservoirs 2310 to perform, forexample, purge/prime cycles.

FIGS. 29A-29B show stages of a purge/prime cycle in accordance with anembodiment of the present technology. Generally, sets of head assemblies2018 can sequentially dispense liquid directly into the reservoirs 2310.When the tray 2050 is positioned to expose about half of the reservoirs2310, one pair of head assemblies 2018 can dispense liquid into exposedreservoirs 2310. The tray 2050 can be moved to expose the other half ofthe reservoirs 2310. Another pair of head assemblies 2018 can dispenseliquid into those exposed reservoirs 2310. The head assemblies 2018 cansequentially dispense liquid while the tray 2050 is positioned withinthe stainer module 2010 to minimize, limit, or avoid off-line times,excessive handling, and/or tray handoffs. As such, a high level ofthroughput can be maintained, even if a large number of purge/primecycles are performed. Additionally, transport problems caused byrepeatedly transporting trays into and out of the stainer module can beavoided. In a purging process, the reservoirs 2310 can collect streamsof liquid from the head assemblies 2018 produced when liquid is pumpedthrough the dispenser heads 2141 to clean any air bubbles from internalpassages. In a priming process, the reservoirs 2310 can collect anyexcess liquid when overfilling the head assemblies 2018 with processingliquid to be dispensed. After performing the purge/prime process, thetray 2050 can be returned to the slide processing position to positionslides underneath each of the head assemblies.

FIGS. 29A and 29B show the tray 2050 at a slide processing position.Referring to FIG. 29B, processing liquid can be delivered onto fourslides while the tray 2050 obstructs a set of vertical delivery paths2380 from the head assembly 2018 d and obstructs a set of verticaldelivery paths 2382 from the head assembly 2018 c. Although the deliverypaths 2380 are illustrated as a single dashed line, each delivery path2380 can extend from one nozzle of the head assemblies 2018 to one ofthe reservoirs 2310. The tray 2050 can collect dispensed liquid that isnot collected by the slides. For example, the tray 2050 can catch liquidthat falls from the slides or drops that fall from the head assemblies2018 (e.g., drops that fall while the tray 2050 is moved to index theslides).

The tray 2050 can be moved from the slide processing position (FIGS. 29Aand 29B) to a purge/prime position 2404 (FIGS. 30A and 30B) forunobstructing the delivery paths 2380. The head assemblies 2018 d, 2018a (head assembly 2018 a is behind the head assembly 2018 d in FIG. 30B)can output liquid along the unobstructed delivery paths 2380. Thereservoirs 2310 a (FIG. 30A), 2310 d can collect the liquid. The tray2050 can be moved from the purge/prime position 2404 (FIGS. 30A and 30B)to another purge/prime position 2410 (FIGS. 31A and 31B) forunobstructing the set of vertical delivery paths 2382. A purge/primecycle can be performed by the head assemblies 2018 b, 2018 c (headassemblies 2018 b is behind head assembly 2018 c in FIG. 31B). In someembodiments, streams of processing liquid are delivered along thevertical delivery paths 2382.

Selected Examples of Liquid Dispensing in Stainers

FIG. 32 is an isometric view of a dispenser apparatus 3024 in accordancewith an embodiment of the present technology. The dispenser apparatus3024 can provide valve-controlled, pressurized liquid delivery andcontrolled movements of head or manifold assemblies 3018 a, 3018 b, 3018c, 3018 d (collectively “head assembles 3018”). A liquid handling system3013 can deliver liquid to the dispenser apparatus 3024 and can include,without limitation, a liquid source 3014 and a liquid conveyance system3015 including conduits or other suitable liquid conveyance elements. Acontroller 3017 can command the dispenser apparatus 3024 to dispense anddistribute protocol-driven liquids over processing zones (i.e., specimenstaining areas of a tray). Controlled liquid distribution can beachieved by, for example, wetting label areas of microscope slides withflowable hydrophobic substances (e.g., to create barriers on label areasof the slides), employing specific liquid exit velocities (e.g.,non-dribbling liquid exit velocities), dispensing specific volumes ofliquid (e.g., volumes of liquid less than an upper volume limit), movinga tray at target speeds and accelerations, dispensing at appropriatedispense locations along the slides, and/or dispensing from targetdispense heights (e.g., heights suitable to minimize or limit splashing,splattering, bouncing of liquid, etc.).

The head assembly 3018 a can move in a direction substantially parallelto a longitudinal axis 3021 of the slide 3020 held by a tray (not shown)while the head assembly 3018 a dispenses liquid. FIG. 33 is a side viewof the head assembly 3018 a with a liquid dispense mechanism 3019(“dispenser mechanism 3019”) dispensing liquid 3030 to form anopen-thick film that covers a specimen 3034 located on an upper surface3044 of the slide 3020. The dispenser mechanism 3019 can include adispenser head 3141, arrays of nozzles 3052, 3054, and shared manifoldswithin the dispenser head 3141. A line 3059 of the liquid conveyancesystem 3015 can deliver liquid from a liquid source 3058 (e.g., multiplecontainers respectively carrying processing liquids) to the dispensermechanism 3019. The liquid flows through the dispenser head 3141 andexits via the nozzles 3052. FIG. 33A shows a flat or non-beveled end ofthe nozzle 3052 through which the stream of liquid flows. A line 3063 ofFIG. 33 can deliver liquid from a liquid source 3062 to the dispensermechanism 3019. The liquid flows through the dispenser head 3141 andexits via the nozzles 3054. In some embodiments, the dispenser head 3141has an internal shared manifold comprised of two separate manifolds,each shared by multiple liquids to isolate incompatible liquids toprevent undesirable liquid interactions. In one embodiment, one manifoldshares up to four compatible liquids that are sequentially dispensed viathe nozzles 3052 and the other manifold shares up to four compatibleliquids that are sequentially dispensed via the nozzles 3054.

FIGS. 34 and 35 are isometric and bottom views, respectively, of thehead assembly 3018 a. The description of the array of nozzles 3052applies equally to the array of nozzles 3054, except as indicatedotherwise. Referring to FIG. 35, the array of nozzles 3052 can be a rowthat spans widthwise (slide edge to slide edge) relative to specimenlocations along the slide 3020 (illustrated in phantom line) such thatthe array of nozzles 3052 is generally aligned with a width W of themicroscope slide 3020. The nozzles 3052 can be evenly or unevenly spacedapart in a direction (indicated by arrow 3060) that is generallyparallel to the slide width W. An angle, if any, defined by thedirection of spacing and the slide width W can be less than about 5degrees, 3 degrees, or 2 degrees. A length L of the array of nozzles3052 can be less than the slide width W such that all of the liquidstreams are directed toward the upper surface 3044 of the slide 3020. Insome embodiments, the array length L is about 70%, 80%, 90%, or 95% ofthe slide width W. However, other array lengths can be used to directstreams of liquid toward an interior region of the slide 3020 to keepthe liquid spaced apart from the edges of the slide 3020. If thedispensed liquid reaches a position near the edges of the slide 3020(e.g., up to about 0.05 inches from the edge of the slide 3020), surfacetension can help keep the liquid from falling off the slide 3020. Thenozzles 3052 are spaced apart in a generally linear arrangement. Inother embodiments, the nozzles 3052 are spaced apart in a U-shapedarrangement, V-shaped arrangement, saw-tooth arrangement (e.g., withdifferent sized nozzles 3052), or other suitable arrangement with anydesired number of nozzles 3052 and any desired nozzle geometries.

FIGS. 36-38 show stages of dispensing liquid in accordance with anembodiment of the present technology. Generally, the dispenser mechanism3019 can deliver liquid at an anti-splatter liquid exit speed tominimize or limit splattering to avoid misprocessing nearbyspecimen-bearing microscope slides. In “painting” dispense processes,the dispenser mechanism 3019 can produce continuous, unbroken lines ofliquid on the upper surface 3044. In “multi-step” dispense processes,liquid can be dispensed in short bursts at specific target locationsalong the slide 3020. The lines of liquid or discrete volumes can spreadout along the upper surface 3044 to cover the specimen 3034 with liquid.In some embodiments, the dispensed liquid can form a matrix of liquidvolumes or lanes of liquid volumes capable of dynamically transforminginto a consolidated film (e.g., a thick film or a puddle). Becausedispense positions relative to the length of the slide 3020 can varybetween liquids, the lengthwise dispense positions can be selected basedon individual liquid characteristics, as well as variability in slidedimensions and slide placement (e.g., placement of slides in the slidecarrier/tray).

FIGS. 36 and 36A show the nozzles 3052 located above a label area of theslide 3020. Referring to FIG. 36A, the nozzles 3052 are orientedvertically to define a flow path 3102, which is substantiallyperpendicular to the upper surface 3044 of the slide 3020. The term“substantially perpendicular” generally refers to an angle within about+/−5 degrees of 90 degrees. For example, an angle α defined by the flowpath 3102 and the upper surface 3044 can be within about +/−5 degrees of90 degrees, such as within about +/−3 degrees of 90 degrees. If theslide 3020 is horizontal, the flow path 3102 can be in a substantiallyvertical orientation. The term “substantially vertical” generally refersto ranges of small angles from vertical, for example, angles betweenabout 0 degrees and 3 degrees from vertical, such as angles less about 2degrees from vertical, for example, angles less than 1 degree fromvertical. The orientation of the nozzles 3052 can be selected based onthe desired liquid interaction with the liquid on the slide 3020. By wayof example, the nozzles 3052 can be at non-vertical orientations to, forexample, produce streams of liquid that push liquid along the slide3020. In some embodiments, the nozzles 3052 can be at substantiallyvertical orientations, and the nozzles 3054 can be at non-verticalorientations.

FIG. 36 shows a slide retainer 3110 (“retainer 3110”) of a tray (notshown) holding the slide 3020. If liquid contacts the retainer 3110, theliquid may tend to wick along the retainer 3110. This wicking may reducethe liquid available for specimen processing and/or cause the formationof undesirable liquid residues. The reductions in liquid available forspecimen processing due to wicking may be relatively imprecise and mayadversely affect the precision of specimen processing. To avoid wicking,the dispenser mechanism 3019 can dispense liquid 3030 a onto the label3026 to form a barrier that keeps subsequently dispensed liquid fromcontacting the retainer 3110. The liquid 3030 a can comprise, withoutlimitation, hydrophobic substances, wax, deparaffinizing liquid, orother suitable substances. The liquid 3030 a can be selected tohydrophobically repel later dispensed aqueous liquids. The barrier canbe temporary, with the residue of the liquid 3030 a eventuallyevaporating. Alternatively, the liquid 3030 a can be selected tosolidify to form a physical barrier after being dispensed.

FIG. 37 shows a barrier 3104 comprised of the liquid 3030 a. The barrier3104 covers an edge 3116 of the label 3026 and can extend along most ofa width W_(L) (FIG. 32) of the label 3026. In some embodiments, thebarrier 3104 extends along a majority of the width W_(L). For example,the barrier 3104 can extend along at least about 70%, 80%, 90%, or 95%of the width W_(L). In one embodiment, the entire label 3026 can becovered by the barrier 3104. The label 3026 can include machine-readablecode (such as a one- or multi-dimensional bar code or infoglyph, an RFIDtag, a Bragg-diffraction grating, a magnetic stripe or a nanobarcode)with coded instructions that specify the type, sequence, and timing ofthe liquid(s) delivered for treatment of a particular specimen. In someembodiments, the label 3026 is a bar code label adhered to the uppersurface 3044.

The dispenser mechanism 3019 can dispense liquid 3030 b (e.g., stainingreagent) at an exit speed (i.e., an anti-splatter exit speed) that isless than a splattering exit speed at which the liquid 3030 b would tendto splatter a liquid film or puddle at least partially supported on theupper surface 3044 by surface tension. In some embodiments, the liquid3030 b is delivered at an anti-splatter exit speed greater than 50cm/second, greater than 57 cm/second, within a range from 50 cm/secondto 60 cm/second, within a range from 54 cm/second to 57 cm/second, aboveanother suitable threshold or within another suitable range. Thecorresponding volumetric flow rate can be, for example, from 0.9 to 1.4mL/second, such as from 1.1 to 1.2 mL/second. In one embodiment, 100 μLto 1500 IA, of liquid 3030 b can be applied to the upper surface 3044 inless than about 5 seconds without any splattering. In some embodiments,100 μL of liquid 3030 b can be delivered onto the upper surface 3044 inless than about 0.1 second, and 1500 μL of the liquid 3030 b can bedelivered onto the upper surface 3044 in less than about 1.5 seconds. Byminimizing or limiting splattering, substantially all of the dispensedliquid 3030 b is collected on the upper surface 3044. For example, atleast about 90% (e.g., at least about 99%) by volume of the dispensedliquid 3030 b can be collected on the upper surface 3044. Thus, lessthan about 10% (e.g., less than about 1%) by volume of the dispensedliquid 3030 b falls into the tray or splatter onto adjacent slides. In aparticular embodiment, from about 99% to about 99.9% or 100% by volumeof the dispensed liquid 3030 b is collected on the upper surface 3044.

Additionally or alternatively, the liquid 3030 b can be delivered at aliquid exit speed greater than a trampoline liquid exit speed. Thetrampoline liquid exit speed is a flow rate at which at least asignificant portion of the stream 3130 would tend to bounce off asurface 3122 of the film or puddle on the slide 3020. The exit speed ofthe stream 3130 can be sufficiently high to avoid trampoline effects butsufficiently low to avoid appreciable splattering. In some embodiments,the liquid 3030 b can exit the nozzles 3052, with inner diameters ofabout 0.24 inch (0.6 mm), at a flow speed in a range about 55 cm/secondto about 60 cm/second. In one embodiment, the liquid 3030 b exits thenozzles 3052 at a flow speed equal to about 57 cm/second. The exit speedof the stream 3130 can be selected based on, for example, the number ofnozzles, nozzle inner diameters, liquid pressures, orientation of thenozzles, height of the nozzles, characteristics of the liquid 3030 b(e.g., viscosity, density, surface tension, etc.), surfacecharacteristics of the slide 3020, and/or environmental characteristics(e.g., surrounding air flow, temperature, humidity, etc.). In someembodiments, at least one nozzle 3052 is spaced apart from the uppersurface of the microscope slide by a distance in a range from about 5 mmto about 10 mm.

FIG. 38 shows the nozzles 3052 dispensing the liquid 3030 b onto an endportion 3150 of the slide 3020 and a film of liquid 3030 b covering mostof a longitudinal length of a processing area 3098 (FIG. 32), such as amounting region or a staining area. The speed of the dispenser mechanism3019, path of the dispenser mechanism 3019, liquid volumetric flow rate,and/or dispense timing can be selected based on the desired liquidcoverage. The dispenser mechanism 3019 can move back and forth along theslide 3020 while dispensing liquid continuously or periodically tomaintain desired coverage. In such processes, the dispensed streams ofliquid can combine with film or puddle upon contact.

The process of FIGS. 36-38 can be used to dispense a wide range ofliquids. By intentionally over-wetting with deparaffinizing liquid (orother oily hydrophobic liquid), a sufficient volume of deparaffinizingliquid can be dispensed at the processing area 3098 (FIG. 32) to provideappropriate liquid spreading to, for example, create the barrier 3104(FIGS. 37 and 38) in order to mitigate unintentional liquid wickingalong the retainer 3110 (FIG. 36). Relatively large deparaffinizingliquid volumes (e.g., 0.92 mL (+11%/−11%)) can be dispensed for initiallabel and tissue area wetting, and the next largest deparaffinizingvolume (e.g., 0.58 mL (+20%/−20%)) can be used for second dispenses(including key deparaffinization dispenses), and relatively smalldeparaffinizing volumes (e.g., 0.44 mL (+59%/−62%)) can be used foradditional deparaffinizing dispenses. Other volumes of deparaffinizingliquids can be dispensed in other sequences. Conditioning liquid (e.g.,transfer or bridging liquid) can be dispensed to maintain a minimumkinetic liquid thickness above the specimen 3034, but the volume ofconditioning liquid can be sufficiently low to prevent spreading to thelabel area, which can affect the barrier 3104. In some embodiments,conditioning liquid comprising di(propylene glycol) propyl ether can bedelivered with a liquid exit speed equal to about 54 cm/second in orderto dispense a volume of about 0.4 mL (+50%/−50%). In some embodiments,washing liquid can be delivered with a liquid exit speed equal to about57 cm/second to dispense a volume of about 1.0 mL (+10%/−10%), 0.9 mL(+22%/−22%), or 1.1 mL (+10%/−10%). The liquid exit speed of stainingreagent (e.g., hematoxylin reagent) can be equal to about 57 cm/secondto dispense a volume of about 1.05 mL (+14/−14%). The liquid exit speedof stain-setting reagent can be equal about 57 cm/second in order todispense a volume of about 1.2 mL (+16/−16%). The liquid exit speed ofcounterstaining reagent (e.g., eosin reagent) can be equal to about 57cm/second in order to dispense a volume of about 1.35 mL (+11/−11%).Other liquid exit speeds can be selected based on, for example, liquidcharacteristics, spacing between slides, target processing volumes,target dispense times, target processing times, and/or other processingparameters.

Dispense locations, both along the slide length and width, can beregistered with respect to particular slide boundaries in order toachieve desired coverage (e.g., full and uniform liquid coverage of theprocessing area) while limiting or preventing unintentional liquidcontact. The width of the head assembly 3018, number of nozzles (e.g.,number of nozzles 3052, number of nozzles 3054, etc.), the spacingbetween nozzles (e.g., spacing between nozzles 3052, 3054), traymovements, and dispense volumes can be selected to accommodate thespreadability of the dispensed volume and positional tolerances impactedby tray handling. In general, both “painting” dispense routines or“multi-step” dispense routines can achieve liquid coverage of the entireprocessing area (or at least about 90%, 95%, or 100% of the area of theprocessing area 3098 of FIG. 32), but painting dispenses may have lesssplash potential than multi-step dispenses. This is because paintingdispenses may have limited breaks in liquid flow between the nozzles anddispensed liquid. Painting dispenses can also reduce or limit processingtimes due to the coordinated relatively high speed movement of the headassemblies 3018. In contrast to painting dispenses which rely on liquidflow rates/valve timing matched to movement of the head assembly 3018,multi-step dispenses can depend on multiple dispenses with relativelyshort valve times and can be generally implemented independent ofmovement speed of the head assembly 3018. For hematoxylin and eosinstaining, both painting and multi-step dispense routines can be used topromote uniform and consistent stain quality. Multi-step dispense andassisted liquid movement (e.g., airknife assisted liquid movement) canenhance rinsing (e.g., rinsing after applying hematoxylin). The totaltime to accomplish dispensing and liquid removal can impact the abilityto achieve a desired overall processing time, as well as the ability tosupport short incubation times (e.g., 2 minutes, 1 minute, 30 seconds,20 seconds, etc.). The dispense process discussed in connection withFIGS. 36-38 can be modified to reduce processing times. For example,specimen 3034 can be processed without utilizing the liquid 3030 adiscussed in connection with FIGS. 36 and 37. The dispenser mechanism3019 can initially dispense liquid 3030 b (FIG. 37) onto a region of theupper surface 3044 spaced apart from the label 3026 to prevent physicalcontact between the label 3026 (or retainer 3110) and the liquid 3030 b.

Referring again to FIG. 32, the controller 3017 can contain instructionsfor commanding the four head assemblies 3018 to process up to fourslides in parallel using, without limitation, label-to-end dispenseroutines (discussed with respect to FIG. 36), end-to-label dispenseroutines, end-to-label-to-middle dispense routines, or other dispenseroutines. In end-to-label dispense routines, liquid can be applied tothe entire length of the slide 3020. In end-to-label-to-middle dispenseroutines, liquid can be delivered while moving the head assembly 3018along the entire length of the slide 3020. After liquid is applied alongthe length of the slide 3020, the head assembly 3018 can be moved backto the middle of the slide while continuing to dispense liquid. Thecontroller 3017 can adjust processing based on one or more signals fromsensors that detect overflow from either a liquid collector (e.g., apurge tray or a purge pan) or slide tray, as well as sensors that candetect, without limitation, inadequate flow velocity (e.g., low flowvelocity due to contamination), nozzle blockages, changes in valvetiming (e.g., valve timing that may affect processing reliability),wicking along slide retainment features (e.g., retainer 3110, clips orposts of a tray, etc.), contamination (e.g., non-nominal slide surfacecontamination), and/or stainer module's inability to remove bubbles/airpockets along flow paths using, for example, dispense routines, such aspurge cycles, prime cycles, combinations thereof (e.g., purge/primecycles). In the event of stainer module 3010 shut down or the detectionof liquid overflow, all liquid dispense valves of the head assemblies3018 can be shut off.

The stainer module 3010 can process trays independent of the slidepositional content within the trays. The controller 3017 can executeinstructions to move the head assemblies 3018 independent of whether aslide is underneath the head assembly 3018. Movements and delays fordispensing and removing liquids can be performed for all slide positionsfor consistent processing between trays. However, the stainer module3010 only dispenses liquid at slide positions at which a slide ispositioned. Thus, processing times for filled trays (i.e., trayscompletely filled with microscope slides) can be the same as processingtimes for partially filled trays.

The dispenser apparatus 3024 can have head assemblies with differentconfigurations. FIGS. 39-48C show one embodiment of the head assembly3018 and its components and functionality. Valves and liquid componentsare generally discussed in connection with FIGS. 39-41. Manifolds andvacuum features are generally discussed in connection with FIGS.42A-48C. FIGS. 49-53 show another head assembly and its components andfunctionality. A person skilled in the relevant art will understand thatthe stainer module 3010 may have other embodiments of head assemblieswithout several of the features described below with reference to FIGS.39-53.

Referring now to FIGS. 39-41 together, the head assembly 3018 caninclude an array of lines 3160 a, 3160 b, 3160 c, 3160 d (collectively“lines 3160”) and an array of lines 3162 a, 3162 b, 3162 c, 3162 d(collectively “lines 3162”). The lines 3160, 3162 can include one ormore flow elements that facilitate controlled liquid dispensing. Suchflow elements can be orifices configured to produce generally uniformliquid pressures within the dispenser head 3141. In some embodiments,the orifices can be configured to induce most of the pressure drop(e.g., at least about 80% of a total pressure drop) along the respectiveline at one location to minimize or limit pressure variation, if any,induced from other system geometry (e.g., tubing lengths, elevation,fittings, valves, etc.) resulting in controlled/low variation liquiddispensing. In one embodiment, the orifices include a jewel orifice anda housing holding the jewel orifice. The jewel orifice can be a rubyorifice having an opening with an inner diameter of about 0.18 inch(0.3046 mm) Other orifices with different configurations and diameterscan also be used.

The head assembly 3018 can include valves 3170 a, 3170 b, 3170 c, 3170 d(collectively “valves 3170”) and valves 3172 a, 3172 b, 3172 c, 3172 d(collectively “valves 3172”) that are staggered to allow increasedrouting density in the dispenser head 3141, but other mountingarrangements can be used. The configurations of the valves 3170, 3172can be selected based on, for example, material compatibility, operatingpressures, target response times, etc. By mounting the valves 3170, 3172directly to the dispenser head 3141, drops caused by “pumping” actionfrom movement of the head assembly 3018 can be reduced or avoided. Thevalves 3170, 3172 can be operated to dispense liquid at appropriate exitvelocities and to prime the nozzles 3052, 3054 prior to dispensingon-slide. Periodic purging/priming cycles can be performed to mitigatenozzle occlusion/plugging caused by, for example, hematoxylinprecipitate or bluing stain salts. In a single liquid dispense state,the head assembly 3018 can dispense processing liquid from only one ofthe lines 3160, 3162. For example, the valve 3170 a can be in an openstate to dispense processing liquid from the line 3160 a while thevalves 3170 b, 3170 c, 3170 d and valves 3172 are in closed states.After dispensing the processing liquid, the valve 3170 a can be switchedfrom the open state to a closed state, and one of the valves 3170 b,3170 c, 3170 d can be switched from the closed state to an open state todispense another liquid. In a mixed liquid dispense state, two or morevalves (e.g., two or more valves 3170 or two or more valves 3172) can bein open states to deliver multiple liquids into a single manifold inwhich the liquids mix. The mixture can flow out of the manifold and thehead assembly 3018. In some stain routines, the head assembly 3018 canswitch between single liquid dispense states and mixed liquid dispensestates.

FIG. 42A is a cross-sectional view of the head assembly 3018 taken alongline 42A-42A of FIG. 41. The head assembly 3018 includes a manifold 3166for distributing liquid from the lines 3160 to the nozzles 3052 and amanifold 3164 for distributing liquid from the lines 3162 to the nozzles3054. Liquids delivered through the lines 3160 may be incompatible withthe liquids delivered through the lines 3162. The two manifolds 3164,3166 can physically separate liquids that have high potential forundesirable interactions. If stain-setting reagent (e.g., bluing) andhematoxylin contact each other, hematoxylin, even at relatively lowconcentrations, can precipitate out and occlude or plug the nozzles. Ifstain-setting reagent contacts certain washing or conditioning liquids,there may be unintended stain artifacts. To avoid these problems,stain-setting reagent can flow through the manifold 3166 and hematoxylinreagent, washing liquid, and conditioning liquid can flow through themanifold 3164. The assignment of liquids (e.g., deparaffinizing liquid,conditioning liquid, washing liquid, and hematoxylin reagent sharing onemanifold while eosin reagent, stain-setting reagent, andstain-differentiating reagent share another manifold) not only keepsappropriate liquids separated from each other but also may allow forefficient liquid exchange. Conditioning liquid, deparaffinizing liquid,washing liquid, and hematoxylin reagent can be delivered through thelines 3162 a, 3162 b, 3162 c, 3162 d, respectively. Stain-settingreagent, eosin reagent, washing liquid (e.g., washing liquid compatiblewith bluing), and stain-differentiating reagent (e.g., acid wash) can bedelivered through the lines 3160 a, 3160 b, 3160 c, 3160 d,respectively. Other assignments of liquids to the lines 3160, 3162 canbe selected based on the compatibility of the liquids in a givenstaining protocol.

FIG. 42B is a detailed view of the manifold 3166. FIG. 43 is across-sectional view of the head assembly 3018 taken along line 43-43 ofFIG. 40. The manifold 3166 can include a distribution chamber 3186,inlets 3188 a-d (collectively “inlets 3188”), and outlets 3189. Eachvalve 3170 can control liquid flow through a respective inlet 3188,which opens into the distribution chamber 3186. The size, shape, andconfiguration of the distribution chamber 3186 can be selected based on,for example, the desired liquid flow through the manifold 3166.Processing liquids can be individually delivered through respectiveinlets 3188 and into the distribution chamber 3186, which in turndistributes the processing liquid to the outlets 3189. The number ofinlets, location of the inlets, and dimensions (e.g., diameters) ofinlets can be selected based on the desired flow through the manifold3166.

Referring now to FIGS. 42A and 42B, the line 3059 c can deliver liquid(represented by arrows) from a liquid source 3058 c to the line 3160 c.The liquid flows through the line 3160 c and proceeds along a valve feedpassageway 3181 c. The valve 3170 c, in an open state, delivers theliquid into a valve outlet passageway 3182 c. Referring now to FIG. 42B,the liquid flows along the valve outlet passageway 3182 c, through theinlet 3188 c, and into the distribution chamber 3186. The liquid flowsthrough the distribution chamber 3186, outlets 3189, and channels 3191and exits via the row of nozzle outlets 3212.

As shown in FIG. 42B, the nozzle 3052 can extend slightly into thedistribution chamber 3186 to mitigate burrs or other features that mayimpede liquid flow and can be made, in whole or in part, of metal (e.g.,stainless steel, aluminum, or the like), plastic, or other materialssuitable for contacting the processing liquids and can have lengths in arange of about 5 mm to about 25 mm. For example, the nozzles 3052 can behollow metal needles. Additionally, the nozzles 3052 can comprise one ormore coatings to enhance performance Inner surfaces and/or outersurfaces of the nozzles 3052 can include a hydrophobic coating to avoidhanging drops. Non-stick coatings (e.g., polytetrafluoroethylenecoatings), low-friction coatings, or other types of coatings can be usedto reduce liquid carryover between dispense cycles. The inner diametersof the nozzles 3052, 3054 can be small enough and the liquid supplypressure high enough to achieve desired exit velocities/flow rates foreach type of liquid. In some embodiments, for example, the innerdiameters of the nozzles 3052, 3054 can be 0.24 inch (0.6 mm), but otherinner diameters can be selected based on desired back pressures.

The nozzles 3052 may have some variation in tolerance due tomanufacturing tolerances that affects where hanging drops tend to form.This is because hanging drops tend to form on the nozzle 3052 with thelargest inner diameter. One of the nozzles 3052 (or a group of nozzles3052) can have slightly larger inner diameters to promote hanging drops,if any, at that larger diameter nozzle 3052. In some embodiments, sixnozzles 3052 can have inner diameters of 0.233 inch+/−0.005 inch (0.69mm+/−0.13 mm) and the inner diameter of another nozzle 3052 can be 0.263inch+/−0.005 inch (0.69 mm+/−0.13 mm) such that the 0.263 inch diameternozzle 3052 will be the largest, even if all the other nozzles 3052 areat extreme ends of their tolerance ranges. The largest inner diameternozzle 3052 will have the least resistance to liquid flow and droplets,if any, will preferentially form on the outlet of that nozzle 3052. Insome embodiments, the largest inner diameter nozzle 3052 can bepositioned and/or oriented to keep hanging drops from falling onto aslide. For example, the largest inner diameter nozzle 3052 can be angledsuch that its outlet 3212 is spaced away from the slide (e.g., to theside of the slide). During high flow periods (e.g., during dispensing),the liquid can impinge upon the upper surface of the slide, but duringlow flows periods, any drops at the outlet 3212 of the largest diameternozzle 3052 will fall without contacting the slide, thus not interferingwith the incubating liquid.

FIG. 44A is a cross-sectional view of the head assembly 3018 taken alongline 44A-44A of FIG. 41. FIG. 44B is a detailed view of the manifold3164. FIG. 45 is a cross-sectional view of the head assembly 3018 takenalong line 45-45 of FIG. 40.

Referring to FIG. 44B, the manifold 3164 can include a distributionchamber 3196, inlets 3198 a-d (collectively “inlets 3198”), and outlets3199. Each valve 3172 can control liquid flow through a respective inlet3198, which opens into the distribution chamber 3196. Referring now toFIGS. 44A and 44B, the line 3162 b delivers liquid (represented byarrows) to a valve feed passageway 3190 b. The liquid proceeds along thevalve feed passageway 3190 b to the valve 3172 b, which in turn deliversthe liquid into a valve outlet passageway 3192 b. The liquid flows intothe distribution chamber 3196 and exits via the nozzles 3054.

FIGS. 46A-46F show stages of operation of the head assembly 3018.Generally, when switching to a new liquid, vacuum valves 3200, 3202 areenergized (i.e., opened) to remove liquid from the manifold 3166. Avalve connected to one of the lines 3160 can be opened to displace theprevious processing liquid and fill the manifold 3166 with new liquid.Dead legs can be minimized and a liquid exchange process can beperformed to inhibit, limit, or substantially eliminatecross-contamination and/or carry-over. To reduce purge/prime volumes forliquid exchanges, the manifolds 3164, 3166 can be configured to provideuniform flow-through of liquids to limit or prevent pockets of low flowvelocities. By dispensing lengthwise on slides and matching the manifoldsizes to the width of the slide, manifold volumes can be furtherreduced. A single purge/prime cycle can generally include (1) a purgeprocess involving drawing a vacuum and/or rinsing of a manifold with thenext liquid to be dispensed and (2) a prime cycle involving dispensingof the next liquid through the manifold and the nozzles. Liquidexchanges can include multiple exchange steps. For example, an exchangefrom hematoxylin to washing liquid can include multiple exchanges (e.g.,three-mini-cycle exchange processes) with wait times to more efficientlyclean the manifold. Stages of sequentially dispensing two liquids arediscussed in connection with FIGS. 46A-46F.

FIG. 46A is a cross-sectional view of the head assembly 3018 taken alongline 46-46 of FIG. 40. Valve 3170 c (FIG. 39) can be in an open state todispense liquid (represented by arrows 3221) from the nozzles 3052. FIG.46B shows the manifold 3166 filled with liquid and all of the valves3170 in a closed state. FIG. 46C shows the liquid being evacuated fromthe distribution chamber 3186 by opening the valves 3200, 3202. Afterevacuating the distribution chamber 3186, the valve 3170 b (FIG. 39) canbe turned on to deliver another liquid into the distribution chamber3196. FIG. 46D shows exchanging of liquid 3221 with another liquidrepresented by dashed line arrows 3222. The liquid 3222 flows toward thevalves 3200, 3202 until the manifold 3166 is completely filled with theliquid 3222, which can also flow through the nozzles 3052. FIG. 46Eshows the nozzles 3052 filled with the liquid 3222 and all of the valves3170 closed. Once the nozzles 3052 are positioned above the slide to beprocessed, the valve 3170 b can be opened to dispense the liquid 3221,as shown in FIG. 46F. The liquid exchange process described inconnection with FIGS. 46A-46F can be performed to dispense liquid fromany one of the lines 3160, 3162.

FIG. 47 is a cross-sectional view of the head assembly 3018 taken alongline 47-47 of FIG. 40. The head assembly 3018 can include a vacuumchamber 3230 in fluid communication with a vacuum source 3240. Thevacuum source 3240 can draw a vacuum via a line 3250 to draw fluid outof the vacuum chamber 3230. In some embodiments, the vacuum source 3240can include, without limitation, one or more pressurization devices,pumps, or other types of devices capable of drawing vacuum pressuregreater than −0.3 psi with a 4 L/min flow rate, although other vacuumpressures and flow rates can be used. In some embodiments, a vacuum candraw liquid away from the outlets of the nozzles to mitigate hangingdrops. Additionally or alternatively, vacuums can be used to removeliquid from the head assembly 3018 to, for example, perform arinse/purge cycle, a calibration routine, etc. The line 3250 caninclude, without limitation, one or more valves (e.g., one-way valves,check valves, etc.), connectors, sensors, orifices, and/or other fluidiccomponents.

FIG. 48A is a cross-sectional view of the head assembly 3018 taken alongline 48-48 of FIG. 41. FIGS. 48B and 48C are detailed views of portionsof the head assembly 3018 in two different evacuation states. In theevacuation state shown in FIG. 48B, the valve 3202 and the valve 3200(FIG. 47) allow liquid flow between the manifold 3166 and the vacuumchamber 3230. The liquid L₁ (represented by arrows) is drawn upwardlythrough the nozzles 3052 and into the distribution chamber 3186. L₁flows through a passageway 3260 and into the valves 3200, 3202, which inturn delivers L₁ into a passageway 3262. L₁ flows through the passageway3262, the vacuum chamber 3230, and the line 3250 (FIG. 47), therebyevacuating the manifold 3166. In the evacuation state shown in FIG. 48C,the valve 3202 has been opened to allow liquid flow between the manifold3164 and the vacuum chamber 3230. Liquid L₂ is drawn upwardly throughthe nozzles 3054 and into the distribution chamber 3196. L₂ flowsthrough a passageway 3264 and into the valve 3202, which in turndelivers the L₂ into the passageway 3262. L₂ flows through thepassageway 3262, the vacuum chamber 3230, and the line 3250 (FIG. 47),thereby evacuating the manifold 3164. In some modes of operation, one ofthe manifolds 3164, 3166 can be empty and maintained under vacuum whilethe other manifold 3164, 3166 is filled with processing liquid. Thus,only one processing liquid will be ready to be dispensed at any giventime. The vacuum can be used to avoid or limit hanging drops or otherproblems that may adversely affect staining.

The dual manifolds 3164, 3166 and vacuum chamber 3230 can help minimizethe complexity and improve reliability of fluidic and wire routingmanagement and also flow characteristic differences between slide trayquadrants and between stainer modules. The manifolds 3164, 3166, theirassociated valves (e.g., valves 3170, 3172), wires, lines (e.g., lines3160, 3162), and fluidic connections can move along slides multipletimes throughout a protocol to consistently distribute liquids alongeach slide, regardless of the slide location. An energy chain bendradius, flexible and material compatible tubing, and a fluidic designcan be selected such that each individual dispense line has the bulk ofthe pressure drop as defined by a precision restrictor orifice, asdiscussed above, and that the shared delivery lines have as littlepressure drop as possible.

FIG. 49 is an isometric view of a head assembly 3300 in accordance withan embodiment of the present technology. FIG. 50 is a top plan view ofthe head assembly 3300. Referring to FIGS. 49 and 50 together, the headassembly 3300 can include a dispenser mechanism 3310 and a liquidremoval device 3320. The dispenser mechanism 3310 includes a dispenserhead 3330 and valves 3340 a, 3340 b, 3340 c, 3340 d (collectively“valves 3340”) positioned in a radial arrangement. The valves 3340control liquid delivery from lines 3350 a, 3350 b, 3350 c, 3350 d(collectively “lines 3350”). A vacuum can be drawn via lines 3362, 3364to evacuate liquid from the dispenser head 3330. Referring now to FIGS.49 and 51, the liquid removal device 3320 has a line 3380 (FIG. 49)fluidically coupled to the nozzle 3387 (FIG. 51) and lines 3382, 3384(FIG. 49) fluidically coupled to a V-shaped airknife 3389 (FIG. 51),respectively. Air delivered via the lines 3382, 3384 (FIG. 49) exits theairknife 3389 (FIG. 51).

FIG. 52 is a cross-sectional view of the head assembly 3300 taken alongline 52-52 of FIG. 50. A manifold 3420 includes an inlet 3412,distribution chamber 3430, and a liquid distributor device 3440. Thedistribution chamber 3430 can have a relatively small volume to minimizeor limit the volume of liquid within the head assembly 3300. The inlets3412 can be circumferentially positioned about the distribution chamber3430 to help equalize the pressure within the distribution chamber 3430.When the valve 3340 b is in an open state, liquid from the line 3350 bflows through passageways 3400, 3410, and the inlet 3412. The liquidflows through the distribution chamber 3430 and the liquid distributordevice 3440 exits via nozzles 3460.

FIG. 53 is an isometric view of the liquid distributor device 3440 inaccordance with an embodiment of the present technology. The liquiddistributor device 3440 can include a bundle of lines 3492 and a flowseparator 3490. In some embodiments, each line 3492 fluidically couplesone manifold outlet 3432 of the flow separator 3490 to one nozzle. Theliquid distributor device 3440 can have other configurations todistribute liquid to other types of nozzles.

FIG. 54 is a cross-sectional view of a nozzle apparatus 3500 inaccordance with an embodiment of the present technology. The nozzleapparatus 3500 can include a main body 3504 and nozzles 3506 coupled themain body 3504. The nozzle apparatus 3500 can be incorporated into thehead assemblies disclosed herein to produce generally uniform flows.Liquid can flow through a main passageway 3510 and nozzle channels 3512(one identified) of the nozzles 3506. In some embodiments, each line3492 can be positioned in one of the channels 3512. However, othercomponents and configurations can be used to dispense liquid.

Selected Examples of Liquid Removal in Stainers

Automated histological systems configured in accordance with at leastsome embodiments of the present technology include stainers havingconfigured to remove dispensed liquid volumes at precisely controlledtimes without displacing the liquid volumes with other liquids. Forexample, a processing head configured in accordance with a particularembodiment of the present technology uses an air knife and an associatedvacuum port to respectively gather and remove dispensed liquid volumes.This manner of dispensing and removing liquid volumes may facilitatewashing and other specimen-processing operations using stationarypuddles or thick films with shapes maintained at least partially bysurface tension. At least partially uncovering a specimen bymanipulating a previously dispensed processing liquid before contactingthe specimen with another processing liquid is expected to enhance theconsistency and controllability of processing times. By way of theory,and not to limit the scope of the present technology, this advantage maybe associated with reducing timing imprecisions associated withimprecise dilution of processing liquids occurring during directliquid-to-liquid exchanges. Alternatively or in addition, washing aspecimen in a stationary pool of liquid may cause residue to be releasedfrom the specimen more evenly and precisely than would occur if thespecimen were washed in a flowing stream of liquid. Other mechanisms arealso possible. Furthermore, the liquid removal features can havedifferent or additional advantages, such as reducing liquid waste.

FIG. 55 is an isometric view of a dispenser apparatus 4024 that includesfour head assemblies 4018 a, 4018 b, 4018 c, 4018 d (collectively “headassembles 4018”). FIGS. 56-58 illustrate stages of a liquid removalprocess performed by the head assembly 4018 a. With reference to FIGS.55-58 together, the head assembly 4018 a can be positioned above amicroscope slide 4020 (“slide 4020”) to dispense liquid onto an uppersurface of the slide 4020. After the liquid has contacted the specimenfor a desired length of time, the head assembly 4018 a can blow theliquid along the slide 4020 (e.g., lengthwise) and draw a partial vacuumto contactlessly remove collected liquid from the slide 4020. Forexample, the head assembly 4018 a can move (as indicated by arrows)relative to the slide 4020 while blowing the liquid and simultaneouslydraw the partial vacuum. Thereafter, additional liquids can besequentially applied to and removed from the slide 4020. In some cases,dispensing a subsequent liquid begins while a previously dispensedliquid is being removed, such as in the same pass of the head assembly4018 a over the length of the slide 4020. In other cases, removal of apreviously dispensed liquid can be complete when dispensing a subsequentliquid begins.

Referring to FIG. 56, a sufficient volume of liquid 4340 can be locatedon the upper surface 4044 of the slide 4020 to maintain a desired liquidvolume (e.g., a kinetic liquid volume) throughout most or all of atarget time period (e.g., a target incubation time) for contact betweenthe specimen and the liquid 4340. The volume of liquid 4340 can providesufficient mass for wholly or incrementally processing the specimen 4034in a desired manner during a predetermined specimen-processing protocol.A flow generator 4352 (e.g., a pump, an air compressor, a blower, a fan,etc.) can pressurize gas (e.g., air, nitrogen, or other gas) that isdelivered to a liquid removal device 4330 of the head assembly 4018 a.The liquid removal device 4330 receives the pressurized gas and producesa gas curtain 4360 (represented by arrows). As the head assembly 4018 amoves (indicated by the arrow 4025) away from an initial position, thegas curtain 4360 pushes the volume of liquid 4340 (e.g., a puddle or athick film of liquid 4340) toward the end 4143 of the stationary slide4020 and can also urge the liquid 4340 toward a suction element 4370 ofthe liquid removal device 4330.

FIG. 57 shows the liquid removal device 4330 at an intermediate positiongenerally midway between the slide ends 4143, 4366. The volume of liquid4340 is contained on a section of the upper surface 4044 in front of thegas curtain 4360 and a section of the mounting area behind the gascurtain 4360 can be substantially free of the liquid 4340.

The suction element 4370 can draw a partial vacuum to suck the liquid4340 from the slide 4020. The head assembly 4018 a can continue to movetoward the slide end 4143 as and/or until the volume of liquid 4340 iscaptivated at the slide end 4143. FIG. 58 shows the volume of liquid4340 captivated by the gas curtain 4360 at an edge 4147 along the slideend 4143. The liquid 4340 can be sucked into the suction element 4370 tolimit the volume of liquid 4340, if any, that falls from the slide 4020into a tray (not shown) carrying the slide 4020. In some embodiments,substantially no liquid 4340 falls from the slide 4020 to keepsubstantially all processing liquids on-slide.

FIGS. 59, 60, and 61 are isometric, bottom, and front views of the headassembly 4018 a in accordance with an embodiment of the presenttechnology. A gas knife 4350 can be V-shaped to partially surround thesuction element 4370. The gas knife 4350 can be used with a variety ofsuitable gases, such as air, nitrogen, air/nitrogen mixtures, or othergases compatible with processing liquids and tissue specimens. As such,although the term “airknife” may be used herein for ease of reference,unless the context clearly indicates otherwise, the term refers to gasknives capable of producing gas curtains comprised of any suitablegases. Thus, the gas knife 4350 can output streams of air (e.g., ambientair, filtered air, etc.) to produce an air curtain, streams of nitrogento produce a nitrogen curtain, or streams of other gases to produceother types of gas curtains.

Referring now to FIG. 59, the gas knife 4350 can include a manifold withside portions 4390 a, 4390 b and a vertex portion 4392. The sideportions 4390 a, 4390 b are generally similar to one another, andaccordingly, the description of one side portion applies equally to theother side portion, unless indicated otherwise. The side portion 4390 acan have a number of holes 4400 a (one identified) selected based on,for example, the desired width of the gas curtain 4360. In someembodiments, the side portion 4390 a has about 10 to about 20 holes. Inone embodiment, including the illustrated embodiment, the side portion4390 a has sixteen linearly arranged holes 4400 a. To produce agenerally uniform substantially V-shaped gas curtain, the holes 4400 a,4400 b (collectively “holes 4400”) can have a generally uniform pitch(i.e., distances between the centers of adjacent holes 4400). To producea non-uniform V-shaped gas curtain, the holes 4400 can be unevenlyspaced apart. Other numbers, patterns, and spacings of the holes 4400can be selected based on the desired configuration and shape of the gascurtain.

Referring now to FIG. 60, an angle β can be defined by the series ofholes 4400 a and the series of holes 4400 b and can be selected based ongeometric factors, such as the width of a corresponding slide surfaceand the geometric relationship between the holes 4400 and the suctionelement 4370. In addition or alternatively, the angle β can be selectedbased on the properties (e.g., viscosity, spreadability, etc.) of theliquid to be collected. In some embodiments, the angle β is in a rangefrom about 80 degrees to about 100 degrees. In one embodiment, the angleβ is about 90 degrees (i.e., 90 degrees+/−3 degrees). In otherembodiments, the angle β is greater than 100 degrees to collect arelatively large volume of relatively low viscosity liquid. In yet otherembodiments, the angle β is less than 80 degrees to collect a smallvolume of relatively high viscosity liquid.

A width W_(h) of the set of holes 4400 is measured in a directiongenerally perpendicular to either the path of travel of the headassembly 4018 a during use or the direction of the longitudinal axis ofthe slide. In some embodiments, the width W_(h) is selected such thatthe gas curtain 4360 extends across the majority of width of the slide4020. For example, the width W_(h) can be equal to or greater than about25 mm, 30 mm, 40 mm, 50 mm, for slides having widths of 25 mm, 30 mm, 40mm, 50 mm, respectively.

The suction element 4370 can be positioned generally along a centerline4413 of the dispenser head 4141 of the head assembly 4018 a. However,the suction element 4370 can be located at other locations, if needed ordesired. The suction element 4370 can include a tubular body 4410 and aninlet port 4412. The tubular body 4410 is spaced apart from the gasknife 4350 such that the inlet port 4412 is positioned directly betweenthe two series of holes 4400 a, 4400 b. In some embodiments, the inlet4112 can be positioned rearwardly of the distal or forward holes 4400 a,4400 b (i.e., the two holes 4400 a, 4400 b identified in FIG. 60), whichproduce leading portions of the gas curtain. The inlet port 4412 canhave a circular opening with a maximum width in a range of about 0.5 mmto about 2 mm, from about 0.5 mm to about 4 mm, from about 3.2 mm toabout 4 mm, or within another suitable range. In other embodiments, theinlet port 4412 can have non-circular openings (e.g., ellipticalopenings, polygonal openings, etc.) to achieve a desired vacuum level.Furthermore, the opening of the inlet port 4412 can be flared orannular.

FIG. 61 shows the suction element 4370 extending downwardly past theholes 4400 a, 4400 b and bottom surface of the gas knife 4350. A line4357 fluidically couples a vacuum source 4353 to the suction element4370. The vacuum source 4353 can include one or more pumps orpressurization devices capable of drawing a partial vacuum such that theflow rate through the suction element 4370 is at or above a target flowrate (e.g., 30 liters/minute, 40 liters/minute, 50 liters/minute). Insome embodiments, the vacuum source 4353 produces a vacuum pressure in arange of about −10 psi to about −0.5 psi (e.g., −2.2 psi+/−0.2 psi) forproducing a flow rate through the suction element 4370 in a range ofabout 37 liters/minute to 50 liters/minute. Other arrangements (e.g.,fluidic systems, vacuum sources, etc.) can be used to provide vacuumpressure to the head assembly 4018 a.

FIGS. 62 and 63 are partial cross-sectional side views of the liquidremoval device 4330 positioned above the slide 4020. An angle α (i.e.,gas knife angle of attack) between the gas curtain 4360 and the uppersurface 4044 of the slide 4020 can be selected based on, withoutlimitation, working pressures, height of the head assembly 4018 a,travel speeds of the head assembly, and/or characteristics of the liquid4340. In some embodiments, the gas curtain 4360 is not perpendicular tothe upper surface 4044 of the slide 4020. For example, the angle α canbe in a range from about 70 degrees to about 80 degrees. In oneembodiment, the angle α is about 4075 degrees (e.g., 75 degrees+/−2degrees) such that the gas curtain 4360 can effectively push the volumeof liquid 4340 along the upper surface 4044 without pushing anappreciable volume of the liquid 4340 off the slide 4020, even whenleading portions of the gas curtain 4360 are moved beyond the distal end4143 of the slide 4020. In one embodiment, the angle α can be about 70degrees (e.g., 70 degrees+/−2 degrees) to enhance pushing of relativelyhigh viscosity liquids. A constant angle α across the width of the slide4020 can be selected to push a volume of liquid with generally uniformproperties. In other embodiments, a varying angle α can be selected topush a volume of liquid with non-uniform properties. For example, aportion of the gas curtain defining a relatively small angle α can bewell suited to push a low viscosity liquid and a portion of the gascurtain defining a relatively large angle cc can be well suited to pusha high viscosity liquid. Other angles of attack can also be used becausethe distribution of residual liquid on the slide 4020 can be largelyinfluenced by the gas knife angle α, as well as movements of the headassembly across the slide 4020 and the height of the head assemblyrelative to a slide 4020.

FIG. 63 shows the suction element 4370 drawing a partial vacuumsufficient to draw the liquid 4340 upwardly through the inlet port 4412without a solid structure of the head assembly contacting the volume ofliquid 4340 and/or the slide 4020. The gas knife pressures can besufficiently low to minimize or limit overwetting and sufficiently highto keep residual volumes at or below a target level. A height H of theinlet port 4412 can be about 0.8 mm to about 3 mm to achieve relativelylow residual volume levels on the slide 4020. In one embodiment, theheight H is in a range of about 1 mm to about 2 mm (i.e., 1 mm to 2mm+/−0.5 mm), but other heights can be selected based on the vacuumlevel. In some embodiments, the flow rate through the suction element4370 can be in a range of about 37 liters/minute to about 50liters/minute at a dynamic pressure between about −1 psi to about −0.2psi, such as about −0.38 to about −0.3 psi for a height H equal to orless than about 3 mm. However, other heights and pressures can be usedbased on the liquid properties that impact fluid dynamics, such asviscosity, surface tension, density, or the like. In some embodiments,the suction element 4370 is configured to produce a vacuum level in arange of about 12 mmHg to about 35 mmHg. The operation of the vacuumsource 4353 can be adjusted to achieve such vacuum levels or otherdesired vacuum levels.

FIGS. 64A-66B illustrate stages of removing the liquid 4340 from theslide 4020. After the liquid 4340 has contacted the specimen 4034 for adesired length of time, the gas knife 4350 can deliver one or morestreams of gas toward the slide 4020 to produce the gas curtain 4360.The gas knife 4350 can be configured to produce the gas curtain 4360using no more than one stream of gas or using two or more streams ofgas. Before, during, and/or after moving the liquid removal device 4330,the head assembly 4018 can contactlessly remove liquid 4340 from theslide 4020 using the suction element 4370. As the gas knife 4350 movesrelative to the slide 4020, for example, the gas curtain 4360 canconfine and move the volume of liquid 4340 away from longitudinal edges4540, 4542 of the slide 4020 while the suction element 4370 draws apartial vacuum to remove the liquid 4340 from the slide 4020 withoutphysically contacting the slide 4020. Different stages of the liquidremoval process are discussed below.

FIG. 64A shows the liquid removal device 4330 and the substantiallyV-shaped gas curtain 4360 positioned along the slide 4020. FIG. 64B is atop plan view of the gas curtain 4360 and the slide 4020. Referring toFIGS. 64A and 64B together, the liquid removal device 4330 is located atan initial position and directs the gas curtain 4360 (shown in FIG. 64B)toward the upper surface 4044. The volume of liquid 4340 can be a film(e.g., a thick-film) or puddle with a shape at least partiallymaintained by surface tension. The gas curtain 4360 in the initialposition can be located along the label 4026. In other embodiments, mostor all of the gas curtain 4360 in the initial position can be locatedbeyond the label end 4366 of the slide 4020 to enable liquid removalfrom the most or all of the label 4026. Vacuum collection, however, canbe delayed to start beyond the retaining features (e.g., slide retainer,clips, clamps, etc.) that hold the label end 4364 of the slide 4020 soas to prevent liquid 4340 from being actively pulled into retainingfeatures of the slide tray. In yet other embodiments, the gas curtain4360 in the initial position can be located along the mounting area ofthe slide 4020, and vacuum collection can begin prior to or after movingthe gas curtain 4360 along the slide 4020.

The gas consumption/flow rate of the gas knife 4350 can be in a range ofabout 8 liters/minute to about 9 liters/minute, for example, about 8.6liters/minute to provide an input gas knife pressure of about 7psi+/−0.2 psi. Excessively high gas knife pressures and/or flow ratescould lead to loss of removed liquid distribution (overwetting) andexcessively low pressures and/or flow rates could lead to residual highresidual volumes. The gas knife 4350 and suction element 4370 cooperateto produce a pressure differential to urge the proximal region 4580 ofthe volume of liquid 4340 away from the longitudinal edges 4540, 4542.In some embodiments, the gas knife 4350 and the suction element 4370produce a low pressure region 4380 (FIG. 64A) at least partiallydefining a collection zone 4550 (illustrated in phantom line) in whichthe liquid 4340 tends to collect. The collection zone 4550 can bepositioned directly below the inlet port 4412 of the suction element4370. For example, the inlet port 4412 can be positioned proximate tothe vertex 4596 (FIG. 64B) of the gas curtain 4360 such that most of thecollection zone 4550 is positioned generally underneath the inlet port4412. The vertex 4596 can be angled or curved.

Referring now to FIG. 64B, the gas curtain 4360 has curtain portions4590, 4592 and a vertex section 4596. The curtain portions 4590, 4592can be positioned along imaginary planes 4594, 4595 that intersect at anangle ω in a range from about 80 degrees to about 100 degrees. Thevertex section 4596 can be moved along a central region 4600 of theupper surface 4044 such that the curtain portions 4590, 4592 confine theproximal region 4580 of the volume of liquid 4340. As the liquid removaldevice 4330 moves lengthwise along the slide 4020, a pressuredifferential can urge the liquid 4340 toward a central longitudinal axis4021 of the slide 4020, as well as the collection zone 4550. In someembodiments, the gas curtain 4360 can be a generally uniform gas curtainthat extends across the width W of the slide 4020. For example, the gascurtain 4360 can be generally uninterrupted and continuous gas curtainthat extends between the longitudinal edges 4540, 4542.

FIGS. 65A and 65B show the liquid removal device 4330 moving along aprocessing path 4551 generally parallel to the longitudinal axis 4021 ofthe slide 4020. The suction element 4370 can provide a partial vacuum toproduce a low pressure region at the collection zone 4550 where thesuction (flow) of the liquid 4340 can occur. A pressure gradient betweenthe low pressure region and the ambient pressure, along with the gasflow interaction between the gas knife 4350 and the suction element4370, can urge the liquid 4340 toward the collection zone 4550. A region4624 of the upper surface 4044 positioned behind the gas curtain 4360can be substantially free of the liquid 4340. There may be, however, asmall volume of residual liquid on the region 4624, but most of thetotal volume of the liquid 4340 on the slide 4020 can be located betweenthe gas curtain 4360 and the end 4143 of the slide 4020. Depending onthe characteristics (e.g., surface tension) of the liquid 4340, most orsubstantially all of the volume of liquid 4340 can be kept in front ofthe gas curtain 4360 moving along processing path 4551. As the gascurtain 4360 advances distally, the liquid 4340 tends to flow along thecurtain portions 4590, 4592, respectively, as indicated by arrows 4629,and the curtain portions 4590, 4592 can urge outer portions 4620, 4622(FIG. 65B) of the puddle of liquid 4340 away from the longitudinal edges4540, 4542, respectively, to reduce the likelihood of liquid 4340falling off the slide 4020. Advantageously, the lengthwise movement andposition of the gas curtain 4360 allows the head assembly 4018 to bemoved at relatively high speeds while keeping the volume of liquid 4340on the slide 4020.

The gas knife 4350 and suction element 4370 can be aligned with theslide 4020 such that the liquid 4340 is effectively directed by the gascurtain 4360 toward the suction element 4370 because widthwise positionof the liquid removal device 4330 relative to slide edges 4540, 4542 canimpact residual volume and residual liquid distribution. As the liquidremoval device 4330 moves relative to the slide 4020, the collectionzone 4550 can be positioned generally along the central region 4600 ofthe upper surface 4044. In some embodiments, the gas curtain 4360 andthe collection zone 4550 can be centrally aligned above the slide 20within +/−0.05 inch (1.27 mm) of the longitudinal axis 4021. If thesuction element 4370 is not close enough to the upper surface 4044,higher residual volumes can also result. As such, the position of thecollection zone 4500, height of the gas knife 4350, height of thesuction element 4370 can be selected to achieve desired liquid removal(including amount and distribution of residual volumes).

FIG. 66A shows the liquid removal device 4330 positioned at the end 4143of the slide 4020. FIG. 66B is a top plan view of the gas curtain 4360of FIG. 66A. The curtain portions 4590, 4592 can extend outwardly pastthe longitudinal edges 4540, 4542, respectively, and the edge 4147 suchthat the volume of liquid 4340 is contained by the gas curtain 4360 andthe slide edge 4147. The gas knife 4350 and vacuum can be turned offwhen the suction element 4370 reaches the end 4143 of the slide 4020 toprevent liquid overwetting to the back side of the slide 4020 and, insome embodiments, also leaves a residual droplet (e.g., a small residualdroplet at location along the upper surface 4044 underneath the suctionelement 4370). The size of the droplet can be minimized or limited bythe configuration of gas knife 4350 and vacuum characteristics definedby, for example, supply pressures, fluidic design, the geometricalrelationship between the gas knife 4350 and the vacuum, and/or theheight of the suction element 4370 and gas knife 4350 with respect tothe upper surface 4044. If the suction element 4370 is too faroff/beyond the upper surface 4044 (or off the edge 4147), liquid capturecan be affected and, in some embodiments, may lead to higher residualvolumes and potential for overwetting. Thus, the end position of thesuction element 4370 can be selected to achieve desired liquid removalwhile residual volumes and/or overwetting.

To minimize or limit the gap between the inlet port 4412 and the uppersurface 4044 of the slide 4020 at the distal end 4143 of the slide 4020,a fixed nominal vertical (e.g., Z-axis) slope is designed into the gasknife assisted vacuum movement axis, bringing the suction element 4370closer to the upper surface 4044 at the slide end 4143 than at the labelend 4366 to achieve relatively small gaps while preventing interferencesbetween the head assembly 4018 and slide and tray features. In someembodiments, the height of the suction element 4370 at the end 4143 canbe equal to or less than about 2 mm+1 mm/−0.5 mm.

The liquid removal process of FIGS. 64A-66B can be performed to removemost or substantially all the volume of liquid 4340. In someembodiments, the liquid removal device 4330 can remove at least 90% ofthe volume of liquid on the upper surface 4044. In other embodiments,the suction element 4370 and gas knife 4350 are configured to cooperateto remove at least 95%, 98%, or 99% by volume of liquid 4340 from theupper surface 4044. Additionally or alternatively, the liquid removalprocess can be controlled based on target maximum residual volumes. Insome embodiments, the liquid removal device 4330 can remove a sufficientvolume of the liquid 4340 such that a maximum residual volume on theupper surface 4044 after liquid removal is less than the maximumresidual volume. In one process, the volume of liquid 4340 on the uppersurface 4044 can be about 0.5 mL to about 0.9 mL of processing liquid,and liquid removal device 4330 can remove a sufficient volume of liquidsuch that the maximum residual volume of liquid 4340 on upper surface4044 is equal to or less than about 50 μL. The liquid removal device4330 can also remove other volumes of liquid to keep the maximumresidual volume of liquid on the slide 4020 at or below an acceptablevolume, such as 30 μL for deparaffinizating liquids, conditioningliquids (e.g., bridging liquids), washing liquids, andstain-differentiating reagents, 20 μL for staining reagents (e.g.,hematoxylin reagents), counterstaining reagents (e.g., eosin reagents),and stain-setting reagents (e.g., bluing), and 10 μL to limit or preventinterference with subsequent processing. For example, the maximumresidual volume of conditioning liquid can be kept sufficiently low toprevent interference with subsequent coverslipping, enhance handability,meet archivalability requirements, and/or limit the release ofundesirable fumes.

In some embodiments, the gas knife 4350 and suction element 4370cooperate to draw the liquid 4340 from the slide 4020 while keeping atotal volume of the liquid, if any, that falls off the slide 4020 equalto or less than a maximum fall-off volume. The fall-off volume can beequal to about 5%, 3%, or 2% by volume of a total volume of liquid 4340on the slide 4020 prior the beginning the liquid removal process. Assuch, the gas knife 4350 and suction element 4370 can be configured tocooperate to draw at least about 95%, 97%, or 98% of the free-standingvolume of the liquid 4340 (i.e., liquid 4340 located along the surface4044 and not incorporated into the specimen 4034) into the suctionelement 4370.

FIGS. 67-70 illustrate stages of removing the liquid 4340 and dispensinganother liquid 4652. Generally, the head assembly 4018 a can move and/orremove at least portion of the volume of liquid 4340 from the specimen4034 so as to at least partially uncover the specimen 4034. The headassembly 4018 a can dispense liquid 4652 that contacts the uncoveredspecimen 4034. This process can be repeated to sequentially remove anddispense any number of liquids. FIG. 67 shows the specimen-bearing slide4020 after the head assembly 4018 a has begun uncovering the specimen4034. Once the nozzles 4052 or nozzles 4054 of the dispenser mechanism4019 are positioned above the slide 4020, another liquid can bedispensed. The head assembly 4018 a can move along the slide 4020 tofurther uncover the specimen 4034. FIG. 68 shows the specimen-bearingslide 4030 after most of the specimen 4034 has been uncovered. Thenozzles 4052 are dispensing liquid 4652 onto the mounting area of theslide 4020 located behind the gas curtain 4360, which serves as abarrier to prevent contact between the volume of the liquid 4340 and thevolume of the liquid 4652. As the head assembly 4018 a moves andcontinuously or intermittently dispenses liquid 4652 along the slide4020, the liquid removal device 4330 can continuously or intermittentlyremove the liquid 4340 from the slide 4020. FIG. 69 shows the liquid4652 contacting the specimen 4034 and the suction element 4370 drawingthe liquid 4340 from the end 4143 of the slide 4020. As the headassembly 4018 a continues to move past the end 4143 of the slide 4020,the dispenser mechanism 4019 can deliver the liquid 4652 to cover thedesired length of the mounting area of the slide 4020. FIG. 70 shows thenozzles 4052 positioned generally above the end 4143 of the slide 4020.Most or all of the mounting area of the upper surface 4044 can becovered by the volume of liquid 4340.

The removal and dispense process of FIGS. 67-70 can be used to removethe bulk of staining, counterstaining, and stain-setting reagentswithout displacing these reagents with continuously flowing washingliquid. This may improve stain quality, counterstain quality, staincontrollability, counterstain controllability, and/or other aspects ofspecimen processing. For example, when the bulk of staining,counterstaining, and stain-setting reagents are displaced withcontinuously flowing washing liquids, minor changes in the attributes(e.g., stain intensity, counterstain intensity, stain hue, and/orcounterstain hue) of specimens may occur. Although often subtle,attribute changes during exchange periods may tend to be impreciseand/or irregular. Therefore, reducing or eliminating attribute changesduring exchange periods can be desirable. In at least some cases,removing the bulk of staining, counterstaining, and stain-settingreagents with gas and then removing residual volumes of these reagentswith generally stationary volumes of washing liquid is expected toreduce or eliminate attribute changes during exchange periods.

FIG. 71 is an isometric view of a liquid removal device 4700 with alinear (e.g., uniplanar) gas knife 4710 and a suction element 4712. Thegas knife 4710 has a plurality of spaced apart holes configured togenerate a gas curtain 4720. The illustrated linear gas curtain 4720extends across the width W of the slide 4020 such that the gas curtain4720 extends past the edge 4540 of the slide 4020. The gas knife 4710can be moved along a processing path 4729 that is generally parallel toa center line or central longitudinal axis 4021 of the slide 4020. Theadvancing gas curtain 4720 can tend to urge a volume of liquid 4740toward a low pressure collection zone 4742 (illustrated in phantomline), which is positioned proximate to the edge 4542 of the slide 4020.The suction element 4712 has an inlet nozzle 4750 with an inlet port4752 positioned to draw in liquid (e.g., collective liquid) at thecollection zone 4742.

FIG. 72 shows the liquid removal device 4700 pushing the liquid 4740along the slide 4020 while the suction element 4712 sucks in liquid4740. As the liquid removal device 4700 advances distally toward the end4143 of the slide 4020, the liquid 4740 tends to flow along the gascurtain 4720 as indicated by arrow 4770. As such, the liquid 4740 can bemoved away from the longitudinal edge 4540. If the liquid 4740 reachesthe edge 4147, surface tension can help keep the liquid 4740 on theupper surface 4044 of the slide 4020.

FIG. 73 shows the inlet port 4752 positioned at a corner 4780 of theslide 4020. The liquid 4740 is captured between the edges 4147, 4542. Insome embodiments, the height of inlet port 4752 can decrease as itapproaches the corner 4780 to help pick up the volume of liquid 4740.

Liquid removal devices of the present technology can have a wide rangeof different types of outlets and gas knives. FIG. 74 is a bottom viewof a liquid removal device 4800 including a nonlinear (e.g., multiplanaror nonplanar) gas knife 4810 and a suction element 4814 in accordancewith an embodiment of the present technology. The gas knife 4810 can beV-shaped and has a series of elongated slots 4820 through which gasflows to produce a gas curtain. In other embodiments, the gas knife 4810can be a U-shaped gas knife. The dimensions (e.g., lengths, widths,etc.) of the elongated slots 4820 can be selected to achieve a desiredgas curtain. The liquid removal devices can have any number of gasknives of different configurations, including V-shape configurations,U-shape configurations, linear configurations. FIG. 75 is a bottom viewof a liquid removal device 4840 with two gas knives 4842, 4844 inaccordance with an embodiment of the present technology. A suctionelement 4852 is located between the gas knives 4842, 4844, and a suctionelement 4845 is proximate to a vertex 4846 of the gas knife 4844. Inoperation, the leading gas knife 4844 and suction element 4845 cancooperate to remove most of a volume of liquid on a microscope slide.Residual volumes of liquid can be subsequently removed using thetrailing gas knife 4842 and suction element 4852.

The liquid removal devices discloses herein can include a plurality ofsuction elements capable of simultaneously or sequentially removingliquid from a microscope slide. By way of example, a plurality ofsuction elements can be positioned between the sides of a gas curtain.The number, position, and spacing of the suction elements can beselected based on the configuration of the gas curtain. For example, twosuction elements can be used with a W-shaped gas knife that produces aW-shaped gas curtain. Other numbers of suction elements can be utilizedfor gas knives having other configurations.

FIG. 76 is an isometric view of two gas knives 4910, 4912 in accordancewith an embodiment of the present technology. FIGS. 77 and 78 are sideviews of the two gas knives 4910, 4912. Referring now to FIG. 76, thegas knives 4910, 4912 are spaced apart to produce gas curtains 4920,4922, respectively, that define a containment gap 4914 for holding avolume of liquid 4916 (e.g., a film, a puddle, etc.). The gas knives4910, 4912 can move together to translate the volume of liquid 4916along the slide 4020. For example, the gas knives 4910, 4912 can moveback and forth to translate the volume of liquid 4916 across one or morespecimens 4930 (one identified in FIGS. 77 and 78). FIG. 77 shows thevolume of liquid 4916 partially covering one specimen 4930, and FIG. 78shows the volume of liquid 4916 covering three specimens 4930. Thedistance D between the gas knives 4910, 4912 can be increased ordecreased to increase or decrease the size of the gap 4914.

The specific embodiments of the dispenser apparatuses and its featureshave been described herein for purposes of illustration, but variousfeatures have not been described for clarity and numerous modificationsmay be made without deviating from the disclosure. The head assemblies,liquid removal devices, and their components configured in accordancewith embodiments of the present technology can be used with a variety ofvacuum systems, pressurized gas systems, and stainer modules. Forexample, the liquid removal device 4700 discussed in connection withFIGS. 71-73, liquid removal devices 4800, 4840 discussed in connectionwith FIGS. 74 and 75, and gas knives 4910, 4912 discussed in connectionwith FIGS. 76-78 can be incorporated into a wide range of differenttypes of head assemblies and in fluid communication with different typesof vacuum systems/pressurized gas systems, etc.

Selected Examples of Thermal Management in Stainers

Implementing enhanced consistency and controllability of processingtemperature in an automated histological staining system can betechnically challenging for a number of reasons. First, the temperaturein a typical histology laboratory typically varies over time due tocycling of heating and air-conditioning equipment and/or other factors.Second, automated histological staining systems are often located nearother equipment (e.g., autoclaves, hoods, etc.) that inconsistentlycause local heating and/or cooling. Third, temperature sensitivitiesamong the diverse components of an automated histological stainingsystem and among the diverse operations performed within an automatedhistological staining system can vary significantly. As anotherconsideration, the processing liquids used in conventional automatedhistological staining systems tend to be highly volatile and, therefore,may evaporate at an unacceptably high rate at high temperatures.Evaporation is generally undesirable because it tends to be associatedwith inconsistent evaporative cooling of specimens duringtemperature-dependent processing, premature drying of specimens andassociated drying artifacts, noxious odors, and heightened explosionrisks, among other issues. Inconsistent evaporative cooling,furthermore, may be proportionally more problematic at high temperaturesthan at low temperatures since wet bulb depression increasesproportionally with dry bulb temperature at constant relative humidity.Issues at relatively low temperatures include, among others, poor (e.g.,unacceptably slow) reaction kinetics for at least some stainingreactions.

Given the presence of some or all of the associated technical challengesstated above and/or other technical challenges not stated herein,selecting a strategy for enhancing consistency and controllability ofprocessing temperature in an automated histological staining system isnot trivial. In a system configured in accordance with a particularembodiment of the present technology, this strategy includes heating aninternal environment of a stainer of the system to cause a baseline(e.g., set-point, steady-state, and/or average) temperature of theinternal environment to be within a range of greater than ambienttemperatures. Processing specimens at elevated temperatures rather thanat depressed temperatures can be advantageous, for example, because itcan sufficiently distinguish the processing from ambient thermalvariability (i.e., ambient thermal “noise”) without unduly slowing thekinetics of staining and/or other temperature-dependentspecimen-processing reactions. Processing specimens at elevatedtemperatures can actually improve the kinetics of at least somespecimen-processing reactions and, therefore, may increase systemthroughput. As another potential advantage, maintaining an internalenvironment of a stainer at a baseline temperature within a range ofgreater than ambient temperatures may be achievable via heating withoutaccompanying cooling. Avoiding the complexity, bulk, power consumption,and/or other drawbacks of cooling systems can be a significant benefit.In embodiments in which specimens are processed at elevatedtemperatures, evaporation and other challenges of processing-liquidcompatibility can be addressed, for example, by the selection ofdifferent (e.g., less volatile) processing liquids. A more detaileddiscussion of this and other aspects of processing liquids used inconjunction with automated histological staining systems configured inaccordance with at least some embodiments of the present technology isprovided below in a separate subsection.

A suitable elevated baseline temperature for specimen processing may beselected as an upper limit of expected ambient temperatures plus asuitable buffer. Sustained temperatures in most histology laboratoryenvironments are expected to fall within a range from 15° C. to 32° C.Equipment commonly located near automated histological staining systemsin these environments is expected to increase the local temperaturearound the systems by from 0° C. to 4° C. in most cases. A suitablebuffer can be, for example, from 1° C. to 14° C. In at least some cases,the reliability of certain components (e.g., valves) within or near astainer of an automated histological staining system may begin todiminish unduly and/or other negative consequences may be associatedwith temperatures over 43° C., 45° C., 50° C., or another suitablethreshold. With these and/or other considerations in mind, specimenprocessing (e.g., staining) in accordance with at least some embodimentsof the present technology is carried out at a baseline temperaturewithin a range from 37° C. to 43° C. In a particular embodiment, abaseline temperature of an internal environment within a stainer duringspecimen processing (e.g., staining) is about 40° C. In otherembodiments, other suitable baseline temperatures can be used, such asother suitable baseline temperatures within a range from 35° C. to 50°C.

Stainers within systems configured in accordance with at least someembodiments of the present technology are internally heated by differenttypes of heaters. For example, a stainer configured in accordance with aparticular embodiment includes one or more heaters that internally heatthe stainer primarily by forced convection and one or more heaters thatinternally heat the stainer primarily by natural convection and/orthermal radiation. These heaters may operate simultaneously ornon-simultaneously. When present, heaters that heat primarily bydifferent respective heating modalities may complement one another. Forexample, a forced-convection heater may be well suited for elevating thetemperature of an internal environment of a stainer to a desiredbaseline temperature relatively quickly, but also prone to promotingundesirable evaporation of processing liquids used within the internalenvironment. In contrast, a heater with a significant mass that isheated conductively and transfers heat to an internal environment of astainer primarily by natural convection and/or thermal radiation mayreach a desired baseline temperature relatively slowly, but may bewell-suited to maintaining the baseline temperature over time withoutpromoting undesirable evaporation of processing liquids used within theinternal environment. Other synergies are also possible.

FIG. 79 is an isometric view of a stainer 5000 configured in accordancewith an embodiment of the present technology. FIGS. 80-82 arecross-sectional views that illustrate components within an internalenvironment 5002 of the stainer 5000. In particular, FIG. 80 is across-sectional side view taken along the line 80-80 in FIG. 79. FIGS.81 and 82 are cross-sectional plan views taken, respectively, along thelines 81-81 and 82-82 in FIG. 80. With reference to FIGS. 79-82together, the stainer 5000 can include a stainer housing 5004 thatdefines the internal environment 5002. In the illustrated embodiment,the stainer 5000 includes a plate 5006 horizontally disposed at anintermediate elevation within the internal environment 5002. The plate5006 can act as a thermal mass with sufficient bulk to modulate theamplitude and/or frequency of transient temperature non-uniformitieswithin the internal environment 5002. For example, the plate 5006 canhave a uniform or non-uniform thickness greater than 0.5 centimeter,such as greater than 1 centimeter. Furthermore, the plate 5006 can bemade of a thermally conductive material, such as aluminum. This mayexpedite heat transfer between the plate 5006 and gas (e.g., air) withinthe internal environment 5002, which may, in turn, expediteequilibration of temperature non-uniformities within the internalenvironment 5002. In other embodiments, the plate 5006 can be replacedor supplemented with a thermal mass having another suitable form,position, and/or composition. In still other embodiments, the stainer5000 can be without a thermal mass.

The plate 5006 can at least partially compartmentalize the internalenvironment 5002 into an upper region 5002 a and a lower region 5002 b.For example, the plate 5006 can occupy at least 50% by area of a planardivision between the upper and lower regions 5002 a, 5002 b.Alternatively, the internal environment 5002 can be uncompartmentalizedor compartmentalized by a compartmentalizing structure other than theplate 5006. The stainer 5000 can include a portal 5008 through which aslide carrier 5009 can be received into the lower region 5002 b. Theportal 5008 can include a door 5010 configured to open by tilting intothe internal environment 5002 rather than by tilting away from theinternal environment 5002. This can be useful, for example, to preventthe door 5010 from obstructing movement of the slide carrier 5009laterally to a handoff position just outside the portal 5008 when thedoor 5010 is open. The portal 5008 can also include a door sensor 5011configured to detect whether the door 5010 is open or closed. Forexample, the door sensor 5011 can include two separate sensors thatrespectively detect the presence of the door 5010 in an openconfiguration and a closed configuration. The door sensor 5011 can beoperably connected to a controller (not shown), which can useinformation from the door sensor 5011 to manage robotic movement of theslide carrier 5009.

Once inside the internal environment 5002, the slide carrier 5009 can besupported within the lower region 5002 b below a pair of openings 5012in the plate 5006. The stainer 5000 can include processing heads 5014(e.g., head assemblies) disposed at least primarily within the upperregion 5002 a. For example, the processing heads 5014 can extend fromthe upper region 5002 a into the lower region 5002 b toward the slidecarrier 5009 through the openings 5012, such as two processing heads5014 through one opening 5012 and another two processing heads 5014through the other opening 5012 or in another suitable arrangement.Alternatively, the processing heads 5014 can be disposed entirely withinthe upper region 5002 a. The plate 5006 can have a first major surface5016 facing downward toward the slide carrier 5009 and a second majorsurface 5018 facing upward. Specimens (not shown) carried by slides 5020(one identified) on the slide carrier 5009 can be relatively near to thefirst major surface 5016 of the plate 5006. For example, the individualslides 5020 can have a major surface on which a specimen is disposed,and the major surfaces of the slides 5020 can be less than 2centimeters, less than 3 centimeters, and/or less than 5 centimetersfrom the first major surface 5016 of the plate 5006. In this vicinity,the temperature modulating effect of the plate 5006 may be stronger thanit is at other portions of the internal environment 5002.

The stainer 5000 can include one or more internal heaters. These heaterscan be individually configured to internally heat the stainer 5000primarily by forced convection, natural convection, thermal radiation,or a combination thereof. For example, the stainer 5000 can include oneor more conductive heating elements 5022 operably coupled to the plate5006. In the illustrated embodiment, the stainer 5000 includes fourconductive heating elements 5022 (individually identified as conductiveheating elements 5022 a-5022 d) operably coupled to laterally spacedapart portions of the plate 5006 along the second major surface 5018 ofthe plate 5006. In other embodiments, the stainer 5000 can includeanother suitable number, type, and/or position of conductive heatingelements 5022 or no conductive heating elements 5022. The conductiveheating elements 5022 can be independently controlled. For example, thestainer 5000 can include temperature sensors (not shown) operablyassociated with respective laterally spaced apart portions of the plate5006. These temperatures sensors can provide input to respectivefeedback control loops that control operation of respective conductiveheating elements 5022. In addition or alternatively, the stainer 5000can include a temperature sensor 5023 configured to measure an airtemperature within the internal environment 5002.

The stainer 5000 can further include one or more forced-convectionheaters 5024. In the illustrated embodiment the stainer 5000 includestwo forced-convection heaters 5024 (individually identified asforced-convection heaters 5024 a and 5024 b) disposed within the lowerregion 5002 b. In other embodiments, the stainer 5000 can includeanother suitable number, type, and/or position of forced-convectionheaters 5024 or no forced-convection heaters 5024. The individualforced-convection heaters 5024 can include a heating element (notshown), a heat sink 5026 operably (e.g., conductively) coupled to theheating element, and a fan 5028 configured to propel gas (e.g., air)over a surface of the heat sink 5026. The heat sinks 5026 can made of athermally conductive material (e.g., aluminum) and can include featureswith relatively high surface area to promote heat transfer to thepropelled gas. For example, the heat sinks 5026 can include,respectively, arrays of upwardly extending cylindrical aluminum whiskers5029 (one identified). The fans 5028 can be laterally spaced apart fromthe slide carrier 5009 and configured to blow gas diagonally upward. Forexample, the fans 5028 can be oriented to have a predominant outputdirection at an angle from 20 degrees to 70 degrees off horizontal, suchas from 30 degrees to 60 degrees off horizontal. Having thisorientation, the fans 5028 may tend to blow gas toward a gap between theslide carrier 5009 and the first major surface 5016 of the plate 5006.In at least some cases, steady movement of gas through this gap mayenhance temperature uniformity within the gap.

FIG. 83 is a flow chart illustrating a method 5100 for operating thestainer 5000 in accordance with an embodiment of the present technology.With reference to FIGS. 79-83 together, the method 5100 can begin withthe stainer 5000 in an inactive state (block 5102). In this state, thestainer 5000 may consume little or no power. From the inactive state,the stainer 5000 can be warmed up (block 5104). Warming up the stainer5000 can include operating the conductive heating elements 5022 and/orthe forced-convection heaters 5024 to achieve a suitable baselinetemperature within the internal environment 5002. In at least somecases, the stainer 5000 is warmed-up while specimens destined forprocessing within the internal environment 5002 undergo processing thatdoes not involve use of the stainer 5000, such as processing within adrying oven (not shown). This may allow the stainer 5000 to be warmed-upwithout delaying processing of the specimens. After the stainer 5000 iswarmed up, if processing of specimens using the stainer 5000 is not yetneeded, the stainer 5000 can be maintained in a standby state (block5106). While in the standby state, the internal environment 5002 can bevacant, but still maintained at a baseline temperature within a range ofgreater than ambient temperatures. In some embodiments, the stainer 5000is maintained in the standby state at all times or nearly all times whena system including the stainer 5000 is powered on and the stainer 5000is not in use. This can be useful, for example, to allow the stainer5000 to have a relatively low wattage allocation while still being readyfor processing specimens on demand in an acceptable time period. When asystem includes multiple stainers 5000 and in other cases, the wattageallocation available for the stainer 5000 may be relatively small, suchas 200 Watts or less.

Processing specimens within the stainer 5000 can begin when the slidecarrier 5009 is introduced into the internal environment 5002 (block5108). Introducing the slide carrier 5009 can include opening the portal5008, moving (e.g., robotically moving) the slide carrier 5009 towardand into the internal environment 5002, and then closing the portal5008. Once inside the internal environment 5002, the specimens can beprocessed (block 5110). A description of specimen processing inaccordance with at least some embodiments of the present technology isprovided below with reference to FIG. 86. In at least some cases, afterthe specimens have been processed, the slide carrier 5009 is held for aperiod of time within the stainer 5000 (block 5112). This may be thecase, for example, when a processing station to which the slide carrier5009 is to be delivered after exiting the stainer 5000 is not yetavailable. When such a processing station becomes available or atanother suitable time, the slide carrier 5009 can be removed from thestainer 5000 (block 5114). Removing the slide carrier 5009 can includeopening the portal 5008, moving (e.g., robotically moving) the slidecarrier 5009 out of the internal environment 5002, and then closing theportal 5008. Thereafter, the method 5100 can include determining whetherthe stainer 5000 should be shut down. If not, the stainer 5000 can beput back into the standby state until needed for processing additionalspecimens.

During all or a suitable portion of the method 5100, the stainer 5000can be internally heated, such as by operating the conductive heatingelements 5022 and/or the forced-convection heaters 5024. This can causean average temperature within the internal environment 5002 to begreater than an ambient temperature, such as an average environmentaltemperature around an exterior of the stainer housing 5004 within a mainhousing (not shown) of a system including the stainer 5000. Operation ofthe conductive heating elements 5022 and/or the forced-convectionheaters 5024 can be controlled to manage the temperature within theinternal environment 5002. For example, the conductive heating elements5022 and/or the forced-convection heaters 5024 can be operatedbimodally, progressively, and/or in another suitable manner using one ormore feedback loops. Input to the feedback loops can includemeasurements of air temperature (e.g., from the temperature sensor5023), measurements of solid-material temperatures (e.g., from one ormore temperature sensors connected to the plate 5006), and/ormeasurements of other suitable dynamic characteristics corresponding tooperation of the conductive heating elements 5022 and/or theforced-convection heaters 5024.

In some embodiments, the conductive heating elements 5022 and theforced-convection heaters 5024 operate collectively. In otherembodiments, the conductive heating elements 5022 operate collectivelyand the forced-convection heaters 5024 operate collectivelyindependently from the conductive heating elements 5022. In still otherembodiments, one or more of the individual conductive heating elements5022 operates independently and/or one or more of the individualforced-convection heaters 5024 operates independently. Independentoperation of at least some of the individual conductive heating elements5022 and/or the individual forced-convection heaters 5024 may facilitatemodulation of temperature non-uniformities within the internalenvironment 5002. For example, the individual conductive heatingelements 5022 can be operated asynchronously to at least partiallycompensate for detected temperature non-uniformities between differentlaterally spaced apart portions of the plate 5006. Alternatively or inaddition, the individual conductive heating elements 5022 and theindividual forced-convection heaters 5024 can operate independently insome instances and collectively in other instances. For example, if theair temperature within the internal environment 5002 exceeds a set upperthreshold, the conductive heating elements 5022 and theforced-convection heaters 5024 can all be shut off to prevent thestainer 5000 from overheating. If the measured temperature continues torise beyond another threshold, power to the stainer 5000 can be shutoff. This can be useful, for example, to reduce or eliminate the risk ofthermally damaging specimens within the internal environment 5002.

FIG. 84 is a plot 5200 of average temperature within the internalenvironment 5002 (y-axis) relative to time (x-axis) during the method5100. Similarly, FIG. 85 is a plot 5300 of average airflow velocitywithin the internal environment 5002 (y-axis) relative to time (x-axis)during the method 5100. For simplicity of illustration, the averagetemperature scale, the average airflow velocity scale, and the timescales in FIGS. 84 and 85 are arbitrary. With reference to FIGS. 79-85together, when the stainer 5000 is inactive, the average temperature canbe the same as or near an ambient temperature. During this period, theforced-convection heaters 5024 can be off and the average airflowvelocity can be low. In contrast, during the warm-up period, theforced-convection heaters 5024 can be operated aggressively, the averageairflow velocity can be high, and the average temperature can increase.When the average temperature reaches a suitable baseline temperature forthe standby state, operation of the conductive heating elements 5022 andthe forced-convection heaters 5024 can be controlled based on feedback.A duty cycle or other similar operational parameter of theforced-convection heaters 5024 may be lower when the stainer 5000 is inthe standby state than when the stainer 5000 is warming-up. Accordingly,as shown in FIG. 85, the average airflow velocity when the stainer 5000is in the standby state can be less than it is when the stainer 5000 iswarming up.

Shortly before the door 5010 is opened and the slide carrier 5009 isintroduced into the internal environment 5002, active circulation of gaswithin the internal environment 5002 can be suspended or slowed toreduce heat loss through the portal 5008. For example, theforced-convection heaters 5024 can be turned off or operated at arelatively low level. This can persist until the slide carrier 5009 isfully introduced into the internal environment 5002 and the door 5010 isagain closed. As shown in FIG. 85, the average airflow velocity whilethe slide carrier 5009 is being introduced can be relatively low, suchas less than 0.1 meters per second. Even if the forced-convectionheaters 5024 are off while the slide carrier 5009 is being introduced,natural convection, residual forced convection, and/or other phenomenamay cause the average airflow velocity to be greater than it is when thestainer 5000 is inactive. As shown in FIG. 84, with less heating fromthe forced-convection heaters 5024 and with some heat loss through theportal 5008, the average temperature may decrease while the slidecarrier 5009 is being introduced. Thereafter, while the slide carrier5009 is within the internal environment 5002 and the specimens are beingprocessed, the average temperature can be relatively high, such as about40° C. or another suitable specimen-processing temperature within one ofthe ranges of specimen-processing temperatures discussed above. Thespecimens can be at least substantially in thermal equilibrium with theinternal environment 5002 while they are processed. For example, anaverage temperature difference between the specimens can be less than 3°C. (e.g., less than 2° C.) while the specimens are being processed. Amore detailed breakdown of the average temperature and the averageairflow velocity while the specimens are being processed is providedbelow with reference to FIGS. 87 and 88.

While the specimens are being held within the internal environment 5002after processing, active circulation of gas within the internalenvironment 5002 can be suspended or slowed. For example, theforced-convection heaters 5024 can be turned off or operated at arelatively low level. This can be useful, for example, to reduceunnecessary evaporation of liquid (e.g., conditioning liquid) in whichthe specimens are immersed. While the slide carrier 5009 is beingremoved from the internal environment 5002, the forced-convectionheaters 5024 can remain off or operating at a relatively low level toreduce heat loss through the portal 5008. As shown in FIG. 85, theaverage airflow velocity while the specimens are being held and whilethe slide carrier 5009 is being removed can be relatively low, such asless than 0.1 meters per second. As shown in FIG. 84, with less heatingfrom the forced-convection heaters 5024, the average temperature candecrease while the specimens are being held. Then, with some heat lossthrough the portal 5008, the average temperature can continue todecrease. After the slide carrier 5009 is removed and the portal 5008 isclosed, the average temperature can progress toward the averagetemperature when the stainer 5000 is inactive or the average temperaturewith the stainer 5000 is in the standby state depending on whether thestainer 5000 is needed for processing additional specimens.

FIG. 86 is a flow chart illustrating a specimen-processing method 5400corresponding to the specimen-processing portion of the method 5100(FIG. 83). The method 5400 can include first deparaffinizing thespecimens (block 5402). Next, the specimens can be conditioned a firsttime (block 5404), such as by reducing the hydrophobicity of thespecimens and/or otherwise chemically preparing the specimens forstaining. The specimens can then be subjected to a first washing (block5406). After the first washing, the specimens can be stained (block5408) (e.g., non-immunohistochemically stained) and then subjected to asecond washing (block 5410). In some cases, the stain is thendifferentiated and regressed (block 5412) and the specimens aresubsequently subjected to a third washing (block 5414). After the thirdwashing, or directly after the second washing if no staindifferentiating and regressing is performed, the stain can be set andits hue adjusted (block 5416), such as by bluing or purpling. Thespecimens can then be subjected to a fourth washing (block 5418). Next,the specimens can be counterstained (block 5420) and then subjected to afifth washing (block 5422) that can also serve to differentiate andregress the counterstain. Finally, the specimens can be conditioned asecond time (block 5424), such as by increasing the hydrophobicity ofthe specimens and/or otherwise chemically preparing the specimens forcoverslipping.

FIG. 87 is a plot 5500 of average temperature within the internalenvironment 5002 (y-axis) relative to time (x-axis) during the method5400. Similarly, FIG. 88 is a plot 5600 of average airflow velocitywithin the internal environment 5002 (y-axis) relative to time (x-axis)during the method 5400. For simplicity of illustration, the averagetemperature scale, the average airflow velocity scale, and the timescales in FIGS. 87 and 88 are arbitrary. With reference to FIGS. 79-88together, during the deparaffinizing, the first transfer, and the firstwash, the forced-convection heaters 5024 can be operated aggressively,the average airflow velocity can be relatively high, and the averagetemperature can steadily increase. By the time the first wash iscomplete, the average temperature and the average airflow velocity canstabilize at respective baseline values. If the deparaffinizing, thefirst transfer, and the first wash are not included in the method 5400(e.g., when the method 5400 is based on a “stain only” recipe), thespecimens can be held until the baseline temperature is reached.

During staining, active circulation of gas within the internalenvironment 5002 can be suspended or slowed. For example, theforced-convection heaters 5024 can be turned off or operated at arelatively low level. This can be useful, for example, to reduceunnecessary evaporation of staining liquid in which the specimens areimmersed during relatively long incubations. As shown in FIG. 88, theaverage airflow velocity during staining can be relatively low, such asless than 0.1 meters per second. As shown in FIG. 87, with less heatingfrom the forced-convection heaters 5024, the average temperature candecrease. During the second wash, the average airflow velocity can berelatively high and the average temperature can increase. Thereafter,the average airflow velocity and the average temperature can stabilizeat respective baseline values until the second transfer. During thesecond transfer, operation of the forced-convection heaters 5024 cantransition toward the operation described above with reference to FIGS.84 and 85 while the specimens are being held. For example, during thesecond transfer, the forced-convection heaters 5024 can be turned off oroperated at a relatively low level.

In some embodiments, the average temperature during different portionsof the method 5400 is adjustable to affect the attributes of specimensprocessed using the stainer 5000. For example, the average temperatureimmediately before and/or during staining can be selected to control theintensity of the resulting stain. Similarly, the average temperatureimmediately before and/or during counterstaining can be selected tocontrol the intensity of the resulting counterstain. Alternatively or inaddition, these average temperatures can be selected in conjunction withone another so as to control the color balance of the stained specimens.For example, the average temperature immediately before and/or duringstaining can be selected to be the same as or different than the averagetemperature immediately before and/or during counterstaining. In otherembodiments, the average temperature during different portions of themethod 5400 can be non-adjustable.

Recipes according to which the specimens are processed may have one ormore temperature components. For example, a given recipe may specify anaverage temperature for staining and an average temperature forcounterstaining. When specimens are processed according to the recipe,operation of the conductive heating elements 5022 and theforced-convection heaters 5024 can be controlled to achieve thespecified temperatures. The average temperatures can be calculatedautomatically based on a user's indication of a desired attribute forthe specimens. For example, a user may select from a list of specimenattributes (e.g., levels of stain intensity) and the system maycalculate appropriate temperatures alone or in conjunction withappropriate times necessary for achieving the selected attributes. Theattributes can include, for example, stain intensity, staining hue,counterstain intensity, counterstaining hue, and/or staining colorbalance. In other embodiments, average temperatures can be enteredmanually. As with other suitable operations carried out within thesystem, a controller (not shown) can use processing circuitry (also notshown) to execute computer-readable instructions stored on memory (alsonot shown) in a non-transitory form to control heating and relatedoperations within the stainer 5000.

Selected Examples of Specimen-Processing Liquids

Specimen processing using an automated histological system may includecontacting specimens and a series of liquids. The series of liquids caninclude, for example, a deparaffinizing liquid, a conditioning liquid, astaining reagent, a stain-differentiating reagent, a stain-settingreagent, a counterstaining reagent, a washing liquid, and acoverslipping liquid. With reference to FIG. 86, during deparaffinizing,a paraffin composition in which the specimens are embedded can be atleast partially removed so as to expose the specimens for furtherprocessing. In at least some cases, deparaffinizing includes iterations(e.g., 4, 5, 6, 7, 8, or another suitable number of iterations) ofdispensing a deparaffinizing liquid onto slides respectively carryingthe specimens, allowing the dispensed deparaffinizing liquid to remainin contact with a paraffin composition in which the specimens areembedded for a suitable period of time so as to solubilize a portion ofthe paraffin composition (e.g., while the deparaffinizing liquid is inthe form of a puddle having a shape maintained at least partially bysurface tension), and then removing the dispensed deparaffinizing liquidalong with a solubilized portion of the paraffin composition. The timeduring which the dispensed deparaffinizing liquid is in contact with thespecimens can be, for example, a time within a range from 15 seconds to45 seconds. In a particular example, the time is 30 seconds.Conventional deparaffinizing liquids at least typically include xylene,which has relatively high toxicity and volatility and a relatively lowflash point. Conventional alternatives to xylene include monoterpenes,such as limonene and pinene. Although monoterpenes tend to be less toxicthan xylene, other properties of monoterpenes may be very similar tothose of xylene. For example, monoterpenes may have relatively highvolatilities and relatively low flash points.

Operating stainers of automated histological systems at elevatedbaseline temperatures may preclude or at least complicate the use ofxylene, monoterpenes, and other conventional deparaffinizing liquids,such as by exacerbating problematic evaporation of these deparaffinizingliquids. The elevated baseline temperatures, however, may alsofacilitate the use of different deparaffinizing liquids, such asdeparaffinizing liquids that would be comparatively poor solvents ofparaffin compositions at ambient temperatures. Instead of xylene ormonoterpenes, deparaffinizing liquids selected or formulated inaccordance with at least some embodiments of the present technologyinclude one or more alkanes, such as one or more petroleum distillatealkanes. The toxicities and volatilities of these deparaffinizingliquids can be lower and the flash points of these deparaffinizingliquids can be higher than those of conventional deparaffinizingliquids, such as xylene and monoterpenes. Due to these and/or otherdifferences, deparaffinizing liquids selected or formulated inaccordance with embodiments of the present technology can be relativelywell suited for use in stainers that operate at elevated baselinetemperatures.

In addition to or instead of being relatively well suited for use instainers that operate at elevated baseline temperatures, deparaffinizingliquids selected or formulated in accordance with at least someembodiments of the present technology are well-suited for other uses forwhich xylene, monoterpenes, and other conventional deparaffinizingliquids would be poorly suited. As an example, deparaffinizing liquidsselected or formulated in accordance at least some embodiments of thepresent technology are well-suited for forming hydrophobic barriers onspecimen-bearing surfaces of slides. These hydrophobic barriers can atleast partially block undesirable migration of less hydrophobic (e.g.,hydrophilic) liquids during specimen processing subsequent todeparaffinizing. Forming hydrophobic barriers for reducing wetting oflabels on specimen-bearing surfaces of slides is discussed above withreference to FIGS. 36-38. Other uses for hydrophobic barriers are alsopossible.

Deparaffinizing liquids selected or formulated in accordance with atleast some embodiments of the present technology have a C9-C18 alkaneconcentration greater than 50% by volume, such as a C10-C16 alkaneconcentration greater than 50% by volume. The alkane concentration caninclude a single alkane or multiple alkanes. Furthermore, the alkanescan be linear, branched, cyclic, or another suitable form.Deparaffinizing liquids selected or formulated in accordance with atleast some embodiments of the present technology have a C14-C16 alkaneconcentration from 10% to 30% by volume and a C9-C15 alkaneconcentration from 70% to 90% by volume. For example, a deparaffinizingliquid selected or formulated in accordance with a particular embodimentof the present technology includes 20% by volume C14-C16 alkanepetroleum distillate and 80% by volume C9-C15 alkane petroleumdistillate. Suitable C14-C16 alkane petroleum distillates include, forexample, Linpar® 1416V available from Sasol Limited (Johannesburg, SouthAfrica). Suitable C9-C15 alkane petroleum distillates include, forexample, Drakesol® 165AT available from Calumet Specialty ProductsPartners, L.P. (Indianapolis, Ind.). The flash points of these and otherdeparaffinizing liquids selected or formulated in accordance withembodiments of the present technology can be greater than 80° C., suchas greater than 100° C.

Instead of being completely free of terpenes, deparaffinizing liquidsselected or formulated in accordance with some embodiments of thepresent technology include a monoterpene (e.g., limonene or pinene) oranother suitable terpene together with a less volatile component. Theterpene, for example, can be well suited for dissolving paraffin and theless volatile component can be well suited for forming a hydrophobicbarrier. Examples of suitable less volatile components include lipids,such as vegetable oils (e.g., peanut oil). A deparaffinizing liquidselected or formulated in accordance with a particular embodiment of thepresent technology includes 80% limonene and 20% vegetable oil. In atleast some cases, these deparaffinizing liquids may be biodegradable.

After deparaffinizing, the specimens may have a residual hydrophobicitythat would be incompatible with staining. The first conditioning of thespecimens after deparaffinizing can include reducing thishydrophobicity. In at least some cases, the first conditioning includesdispensing a conditioning liquid onto the slides, allowing the dispensedconditioning liquid to remain in contact with the specimens for asuitable period of time so as to wholly or incrementally condition thespecimens (e.g., while the conditioning liquid is in the form of apuddle having a shape maintained at least partially by surface tension),and then removing the dispensed conditioning liquid. The time duringwhich the dispensed conditioning liquid is in contact with the specimenscan be, for example, a time within a range from 5 seconds to 15 seconds.In a particular example, the time is 10 seconds. The conditioning liquidcan be a liquid that is soluble in both a hydrophobic deparaffinizingliquid and water.

Conventional methods for conditioning specimens after deparaffinizingand before staining at least typically include contacting specimens withanhydrous ethanol and then with graded ethanol and water mixtures havingdecreasing concentrations of ethanol and increasing concentrations ofwater. For example, a conventional method may include contactingspecimens with anhydrous ethanol, then a mixture of 95% ethanol and 5%water, and then a mixture of 90% ethanol and 10% water. The initialcontact with anhydrous ethanol may serve to displace the deparaffinizingliquid. The subsequent contact with graded ethanol and water mixturesmay serve to prepare the specimens for contact with aqueous solutions.Without the initial contact with anhydrous ethanol, residualdeparaffinizing liquid would likely persist. Without the subsequentcontact with graded ethanol and water mixtures (i.e., if the specimenswere contacted with an aqueous solution directly after being contactedwith anhydrous ethanol), delicate specimens would likely be damaged.

The use of anhydrous ethanol and graded ethanol and water mixtures forconditioning deparaffinized specimens in conventional methods isproblematic for several reasons. Ethanol, like xylene and monoterpenes,has a relatively low flash point and a relatively high volatility. Forthese and/or other reasons, ethanol may be poorly suited for use atelevated baseline temperatures, which tend to exacerbate problematicevaporation. Problematic evaporation of ethanol may even occur atambient temperatures. Furthermore, anhydrous ethanol readily absorbsmoisture from air. For this reason, protocols associated with storageand use of anhydrous ethanol tend to be burdensome. As yet anotherdrawback, separate plumbing and/or other separate components foranhydrous ethanol and for each different graded ethanol and watermixture can appreciably increase the cost, complexity, and/or bulk ofautomated histological systems.

Instead of anhydrous ethanol and graded ethanol and water mixtures,conditioning liquids selected or formulated in accordance with at leastsome embodiments of the present technology include one or more glycolethers, such as one or more propylene-based glycol ethers (e.g.,propylene glycol ethers, di (propylene glycol) ethers, and tri(propylene glycol) ethers, ethylene-based glycol ethers (e.g., ethyleneglycol ethers, di(ethylene glycol) ethers, and tri(ethylene glycol)ethers), and functional analogs thereof. The flash points andvolatilities of these conditioning liquids can be higher and lower,respectively, than those of conventional conditioning liquids, such asethanol and graded ethanol and water mixtures. Due to these and/or otherdifferences, conditioning liquids selected or formulated in accordancewith embodiments of the present technology can be relatively well suitedfor use at elevated baseline temperatures. Furthermore, relative toanhydrous alcohol, conditioning liquids selected or formulated inaccordance with embodiments of the present technology may have longershelf lives and may have few, if any, special storage and userequirements.

In at least some cases, conditioning liquids selected or formulated inaccordance with embodiments of the present technology are configured foruse in a single formulation. For example, in these cases, it may bepossible, without determent, to contact a specimen with one or morevolumes of a single formulation of a conditioning liquid so as todisplace residual quantities of a deparaffinizing liquid (e.g., a C9-C18alkane) and then contact the specimen with an aqueous wash withoutintervening contact between the specimen and a diluted formulation ofthe conditioning liquid. The risk of damage to these specimens may benegligible or at least less than it would be if the specimens werecontacted with the same aqueous solution directly after being contactedwith anhydrous ethanol. Furthermore, the number of operations involvedin conditioning specimens using conditioning liquids selected orformulated in accordance with embodiments of the present technology maybe less than it would be using conventional conditioning liquids. Forexample, conditioning specimens in methods in accordance with at leastsome embodiments of the present technology includes three or feweriterations of dispensing a conditioning liquid onto slides respectivelycarrying the specimens, allowing the dispensed conditioning liquid toremain in contact with the specimens for a suitable period of time so asto wholly or incrementally condition the specimens, and then removingthe dispensed conditioning liquid. A specimen-processing method inaccordance with a particular embodiment of the present technologyincludes two such iterations. In contrast, a typical conventionalspecimen-processing method includes five or more correspondingiterations. The relatively low number of iterations associated withconditioning in specimen-processing methods in accordance with at leastsome embodiments of the present technology can increasespecimen-processing throughput and/or have other benefits.

Conditioning liquids selected or formulated in accordance with at leastsome embodiments of the present technology have greater volumetricconcentrations of polyol than of monohydric alcohol or of water. Forexample, the conditioning liquids can be non-aqueous and can includegreater than 50% by volume glycol ether, such as greater than 50% byvolume di(propylene glycol) ether and/or tri(propylene glycol) ether. Anon-aqueous conditioning liquid selected or formulated in accordancewith a particular embodiment includes at least substantially exclusivelya mixture of di(propylene glycol) methyl ether and di(propylene glycol)propyl ether. A non-aqueous conditioning liquid selected or formulatedin accordance with another embodiment of the present technology includesat least substantially exclusively di(propylene glycol) propyl ether.Suitable glycol ethers include, for example, DOWANOL products availablefrom Dow Chemical Company (Midland, Mich.). These and other conditioningliquids selected or formulated in accordance with embodiments of thepresent technology can have flash points greater than 70° C., such asgreater than 80° C.

After deparaffinizing and conditioning, the first washing can includeiterations (e.g., 2, 3, or another suitable number of iterations) ofdispensing a washing liquid onto the slides, allowing the dispensedwashing liquid to remain in contact with the specimens for a suitableperiod of time so as to wholly or incrementally wash the specimens(e.g., while the washing liquid is in the form of a puddle having ashape maintained at least partially by surface tension), and thenremoving the dispensed washing liquid. The time during which thedispensed washing liquid is in contact with the specimens can be, forexample, a time within a range from 5 seconds to 15 seconds. In aspecimen-processing method in accordance with a particular embodiment ofthe present technology, this time is 10 seconds. Conventionally, puredeionized water is used as a washing liquid. In contrast, washingliquids selected or formulated in accordance with embodiments of thepresent technology can include deionized water along with a solvent. Thesolvent, for example, can be a polyol, such as propylene glycol. Forexample, the washing liquid can include from 40% to 60% by volumepolyol, such as from 40% to 60% by volume propylene glycol. As furtherdiscussed below, the solvent in the washing liquid can be the same as,within the same chemical class as, or otherwise functionally analogousto solvents included in other liquids that contact the specimens afterthe first washing. Including the solvent in the washing liquid can beuseful to condition the specimens for contacting these other liquids. Asdiscussed below, in at least some cases, the washing liquid is used forcounterstain differentiating and regressing in addition to washing. Inthese cases, the solvent concentration in the washing liquid can beselected both to facilitate the performance of the washing liquid forcounterstain differentiating and regressing and to promote compatibilitywith other specimen-processing liquids.

Washing liquids selected or formulated in accordance with at least someembodiments of the present technology include a surfactant to facilitatespreading of the washing liquids over the specimen-bearing surfaces ofthe slides. The surfactant can be selected to have little or no negativeimpact on specimen-processing operations subsequent to the firstwashing. For example, the surfactant can be non-ionic so as to reduce orprevent undesirable buffering. In at least some cases, the surfactantincludes an ethoxylated alcohol and/or a glycol ether. Suitableethoxylated alcohol surfactants include, for example, TOMADOL® 900available from Air Products and Chemicals, Inc. (Allentown, Pa.) andMerpol SH® available from Stepan Company (Northfield, Ill.). Suitableglycol ether surfactants include, for example, TERGITOL® NP-9 availablefrom Dow Chemical Company (Midland, Mich.).

After the first washing, staining the specimens can include dispensing astaining reagent onto the slides, allowing the dispensed stainingreagent to remain in contact with the specimens for a suitable stainingincubation time so as to stain the specimens (e.g., while the stainingreagent is in the form of a puddle having a shape maintained at leastpartially by surface tension), and then removing the dispensed stainingreagent. The staining incubation time can be, for example, within arange from 1 minute to 20 minutes. In a specimen-processing method inaccordance with a particular embodiment of the present technology, thestaining incubation time is 2 minutes. The staining reagent can beselected or formulated to adequately stain nuclear components of thespecimens without causing unacceptable staining of non-nuclearcomponents of the specimens or other forms of unacceptable non-specificbackground staining. The staining reagent can be anon-immunohistochemical staining reagent, such as anon-immunohistochemical staining reagent including hematoxylin/hematein,a mordant, and a solvent. The solvent can serve to maintain hematein andhematein-mordant complexes in solution. In conventional stainingreagents, the solvent is often ethanol. As discussed above inconjunction with the conditioning liquid, use of ethanol in automatedhistological systems, such as automated histological systems includingstainers configured to operate at elevated baseline temperatures, can beproblematic. Furthermore, staining incubations tend to be relativelylong, which may exacerbate the potential negative effect of ethanol'stendency to evaporate rapidly.

Instead of ethanol, staining reagents selected or formulated inaccordance with at least some embodiments of the present technologyinclude a polyol, such as ethylene glycol, propylene glycol, or acombination thereof. For example, the staining reagents can includegreater than 10% by volume polyol, such as from 10% to 40% by volumepolyol. As discussed below, staining reagents selected or formulated inaccordance with at least some embodiments of the present technologyinclude relatively low concentrations of mordant. This can allow for theuse of relatively high concentrations of solvent, such as concentrationsgreater than 20% by volume. In conventional staining reagents withaverage or high mordant concentrations, these concentrations of solventmay prevent the mordant from adequately dissolving.

Variables that can affect the intensity and selectivity of hematoxylinstain include the pH of the staining reagent, the concentration ofmordant in the staining reagent, the concentration ofhematoxylin/hematein in the staining reagent, and the stainingincubation temperature. Independently, the pH of the staining reagent,the concentration of hematoxylin/hematein in the staining reagent, andthe staining incubation temperature tend to be directly proportional tothe rate at which stain intensity increases, while the concentration ofmordant in the staining reagent tends to be inversely proportional tothe rate at which stain intensity increases. In general, the rate atwhich stain intensity increases is inversely proportional to stainingselectivity. Thus, independently, the pH of the staining reagent, theconcentration of hematoxylin/hematein in the staining reagent, and thestaining incubation temperature tend to be inversely proportional tostaining selectivity, while the concentration of mordant in the stainingreagent tends to be directly proportional to staining selectivity. Thesame correlations may also apply to the effect of the pH of the stainingreagent, the concentration of hematoxylin/hematein in the stainingreagent, and the concentration of mordant in the staining reagent onshelf-life.

Greater rates at which stain intensity increases, greater stainingselectivity, and greater shelf life all tend to be desirable properties.For example, greater rates at which stain intensity increases mayenhance specimen-processing throughput, greater shelf life may enhanceconvenience for users, and greater staining selectivity may enhancestain quality. Although the variables that affect these features can beconsidered independently, they may actually be highly interrelated.Attributes of staining reagents selected or formulated in accordancewith embodiments of the present technology may allow the stainingreagents to take advantage of one or more of the interrelationshipsamong these variables to enhance the balance of staining speed, stainingselectivity, and shelf-life. Furthermore, staining reagents selected orformulated in accordance with at least some embodiments of the presenttechnology have properties that facilitate adjusting hue and/orintensity of nuclear staining via time and/or temperature. Thesestaining reagents can be well suited for use in at least some stainershaving temperature-controlled internal environments in automatedhistological systems configured in accordance with embodiments of thepresent technology.

During hematoxylin staining, the stain intensity may increase steadilyuntil equilibrium is reached. At equilibrium, the rate of deposition ofhematein-mordant complexes from the staining reagent onto the specimenand the rate of release of hematein-mordant complexes from the specimeninto the staining reagent may be approximately equal. The stainintensity at equilibrium tends to be highly dependent on thehematoxylin/hematein concentration in the staining reagent. Stainingreagents with relatively low hematoxylin/hematein concentrations mayreach equilibrium at relatively low stain intensities. Thus, thesestaining reagents may not be capable of producing dark stains even afterlong staining incubation times. This, coupled with the conventionalassumption that the low staining incubation times for producing lightstains using staining reagents with relatively high hematoxylin/hemateinconcentrations are too difficult to control, has motivated theconventional use of two or more different formulations ofhematoxylin/hematein staining reagents in order to produce a full rangeof hematoxylin stain intensities. For example, a conventional set ofstaining reagents for producing a full range of hematoxylin stainintensities at least typically includes one or more staining reagentswith relatively high hematoxylin/hematein concentrations for producingdark stains that cannot be produced using staining reagents withrelatively low hematoxylin/hematein concentrations and one or morestaining reagents with relatively low hematoxylin/hemateinconcentrations for producing light stains considered too difficult toproduce using staining reagents with relatively highhematoxylin/hematein concentrations.

Automated histological systems configured in accordance with embodimentsof the present technology and sets of liquids selected or formulated foruse with these systems can be capable of reliably achieving a full rangeof hematoxylin stain intensities using a single hematoxylin stainingreagent formulation. For example, the control over staining incubationtime achievable with these systems may make it possible to reliablyachieve light stains using staining reagents with relatively highhematoxylin/hematein concentrations. Accordingly, staining reagentsselected or formulated in accordance with at least some embodiments ofthe present technology can have relatively high hematoxylin/hemateinconcentrations, such as hematoxylin/hematein concentrations within arange from 5 to 6.5 grams per liter, within a range from 5.75 to 6.3grams per liter, or within another suitable range. In at least somecases, the hematoxylin/hematein concentrations of the staining reagentsare selected to be as high as possible without unacceptably diminishingshelf life due to the formation of precipitate. The staining reagentscan further include sodium iodate or another suitable oxidizing agent tochemically accelerate ripening of hematoxylin into hematein. Theconcentration of sodium iodate in the staining reagents can be, forexample, less than 10% by weight.

Use of staining reagents having relatively high hematoxylin/hemateinconcentrations can advantageously reduce staining incubation times andthereby increase specimen-processing throughput. It is expected thatthis advantage may exist even with respect to staining reagents havingrelatively low pH. Thus, it may be possible to take advantage of theexpected benefit of relatively low pH on staining selectivity withoutunduly sacrificing staining speed. The pH of staining reagents havingrelatively high hematoxylin/hematein concentrations and other stainingreagents selected or formulated in accordance with embodiments of thepresent technology can be, for example, within a range from 2.4 to 2.6,within a range from 2.45 to 2.54, or within another suitable range. Inat least some cases, the pH is selected to be as low as possible withoutrisking unacceptable damage to specimens, such as damage due to acidhydrolysis of lipids within the specimens. These staining reagents canbe buffered or unbuffered. When buffered, the staining reagents caninclude a suitable buffering agent, such as phthalic acid,chloroacetates, sulfates, glycine, and alanine.

Staining reagents selected or formulated in accordance with at leastsome embodiments of the present technology have enhanced sensitivity totemperature. When used in temperature-controlled stainers of automatedhistological systems configured in accordance with at least someembodiments of the present technology, staining incubation temperaturecan be used alone or in conjunction with staining incubation time tocontrol stain intensity. In general, higher temperatures may causestaining speed to increase and staining selectivity to decrease andlower temperatures may cause staining speed to decrease and stainingselectivity to increase. Temperature can also affect stain intensity atequilibrium. In at least some cases, temperature-dependent stainingreagents selected or formulated in accordance with embodiments of thepresent technology have relatively low mordant concentrations. The stainintensity at equilibrium using these staining reagents may besignificantly more sensitive to temperature than the stain intensity atequilibrium using staining reagents having higher mordantconcentrations.

It is expected that staining using a staining reagent having arelatively low mordant concentration can be taken to equilibrium atdifferent staining incubation temperatures to achieve a full range ofstain intensities. Alternatively, staining using these staining reagentscan be stopped before it reaches equilibrium and temperature and timecan be used together to achieve some or all intensities within the fullrange of stain intensities. In at least some cases, staining incubationtemperature and time can be modified readily. Thus, a user may be ableto use a single staining reagent and select temperature to favorstaining speed at the expense of some staining selectivity or to favorstaining selectivity at the expense of some staining speed depending oncircumstances. Suitable concentrations of mordant intemperature-dependent staining reagents selected or formulated inaccordance with embodiments of the present technology can be less than150% (e.g., less than 125% or less than 100%) of the concentration ofhematoxylin/hematein in the staining reagents. The mordant can be analuminum salt, such as aluminum sulfate hydrate. Salts of other metals(e.g., iron, copper, vanadium, molybdenum, tungsten, indium, nickel,zinc, barium, cobalt, and manganese) can be used instead of aluminumsalt to achieve different stain hues and/or selectivities.

Staining reagents selected or formulated in accordance with embodimentsof the present technology can include other suitable components inaddition to solvent, hematoxylin/hematein, buffer, and mordant. Forexample, the staining reagents can include one or more antioxidants.Antioxidants can be useful, for example, to reduce the formation ofprecipitate and thereby extend the shelf life of staining reagents. Whenpresent, suitable antioxidants include, among others, phenolicantioxidants, such as gallic acid and hydroquinone. As another example,the staining reagents can include one or more stabilizers, such asbeta-cyclodextrin or other suitable cyclodextrins. A staining reagentselected or formulated in accordance with a particular embodiment of thepresent technology includes 747 mL of deionized water, 252.7 mL ofethylene glycol, 6.06 grams of hematoxylin, 0.65 grams of sodium iodate,26.67 grams of aluminum sulfate hydrate, 932 grams of hydroquinone, and11.35 grams of beta-cyclodextrin.

After staining, the second washing can be used to remove residualstaining reagent from the specimens and to increase the pH of the liquidcontent of the specimens sufficiently to halt further staining. Thesecond washing can include use of the same washing liquid and protocoldiscussed above for the first washing. After the second washing, staindifferentiating can be performed to at least partially remove stain frommucin and other non-nuclear portions of the specimens. In at least somecases, stain regressing to lighten nuclear staining of the specimensoccurs in conjunction with stain differentiating. Stain differentiatingand regressing can include dispensing a stain-differentiating liquidonto the slides, allowing the dispensed stain-differentiating liquid toremain in contact with the specimens for a suitable period of time so asto cause sufficient stain differentiating and regressing (e.g., whilethe stain-differentiating liquid is in the form of a puddle having ashape maintained at least partially by surface tension), and thenremoving the dispensed stain-differentiating liquid. The time duringwhich the dispensed stain-differentiating liquid is in contact with thespecimens can be, for example, a time within a range from 30 to 120seconds.

The stain-differentiating liquid can be acidic and can include deionizedwater, an acid (e.g., acetic acid), and a solvent. As with the washingliquid and the staining reagent, the solvent can be a polyol, such asethylene glycol, propylene glycol, or a combination thereof. Forexample, the stain-differentiating liquid can include greater than 10%by volume polyol, such as from 10% to 40% by volume polyol. The use ofat least some conventional stain-differentiating liquids, especially inconjunction with relatively long stain-differentiating incubations, maycause morphological damage to structures within specimens. The use of apolyol solvent in stain-differentiating liquids configured in accordancewith at least some embodiments of the present technology may help tocondition these structures against this type of morphological damage. Inaddition or alternatively, stain-differentiating liquids configured inaccordance with embodiments of the present technology can includerelatively low concentrations of acid to further reduce the possibilityof causing morphological damage to structures within specimens. Forexample, the pH of these stain-differentiating liquids can be greaterthan 2.5, such as greater than 2.7. A stain-differentiating liquidselected or formulated in accordance with a particular embodiment of thepresent technology includes about 700 mL deionized water, 4 mL glacialacetic acid, and 250 mL of propylene glycol. The pH of thestain-differentiating liquid can be, for example, within a range from2.9 to 3.1.

In at least some cases, in addition to being used for staindifferentiating and regressing, the stain-differentiating liquid can beused to remove and/or reduce formation of hematoxylin-containingprecipitates within components of automated histological systems. Forexample, in these cases, the stain-differentiating liquid can be flushedthrough lines and other components of the system that ordinarily carrythe staining reagent to remove and/or reduce formation ofhematoxylin-containing precipitates. In addition to or instead of usingthe stain-differentiating liquid, systems configured in accordance withembodiments of the present technology can use one or more other cleaningliquids for this purpose and/or other purposes. A cleaning liquidselected or formulated in accordance with a particular embodiment of thepresent technology includes about 480 mL deionized water, 500 mLpropylene glycol, and 16.67 mL 6N hydrochloric acid. A cleaning liquidselected or formulated in accordance with another embodiment of thepresent technology includes 450 mL deionized water, 500 mL propyleneglycol, 59 grams trisodium citrate dihydrate, and 50 mL 1N hydrochloricacid.

After stain differentiating and regressing, the third washing can beused to remove residual stain-differentiating liquid from the specimens.The third washing can include use of the same washing liquid andprotocol discussed above in the context of the first and secondwashings. After the third washing, stain setting and hue adjusting(e.g., bluing or purpling) can include exposing the specimens to anenvironment that tends to stabilize hematoxylin-mordant-DNA complexesand to change the stain hue. Stain setting and hue adjusting can includedispensing a stain-setting reagent onto the slides, allowing thedispensed stain-setting reagent to remain in contact with the specimensfor a suitable period of time so as to cause sufficient stain settingand hue adjusting (e.g., while the stain-setting reagent is in the formof a puddle having a shape maintained at least partially by surfacetension), and then removing the dispensed stain-setting reagent. Thetime during which the dispensed stain-setting reagent is in contact withthe specimens can be, for example, about 30 seconds. The stain-settingreagent can include an alkaline solution (e.g., a buffered alkalinesolution) and a solvent. As with the washing liquid, the stainingreagent, and the stain-differentiating liquid, the solvent can be apolyol, such as ethylene glycol, propylene glycol, or a combinationthereof. For example, the stain-setting reagent can include greater than10% by volume polyol, such as from 10% to 60% by volume polyol. Astain-setting reagent selected or formulated in accordance with aparticular embodiment of the present technology includes about 700 mLdeionized water, 12.1 grams of tris(hydroxymethyl)aminomethane, 28.4 mLof hydrochloric acid, and 250 mL of propylene glycol.

The pH of the stain-setting reagent can be selected to change the hue ofthe stain. For example, stain-setting reagents having higher pH cancause more rapid progression to a blue color than stain-setting reagentshaving lower pH. Thus, given a set period of time during which specimensare exposed to a stain-setting reagent, if the stain-setting reagent hasa relatively high pH (e.g., greater than 9), the resulting stain may beblue, whereas if the stain-setting reagent has a relatively low pH(e.g., less than 8), the resulting stain may be purple. Furthermore,when the period of time during which specimens are exposed to astain-setting reagent is relatively long and the stain-setting reagenthas a relatively low pH (e.g., less than 8), the temperature duringstain setting and hue adjusting can be used to change stain hue, such asthe relative level of bluing. As discussed above in the context ofchanging temperature to adjust stain intensity, temperature can be moreconvenient to adjust than the properties (e.g., pH) of a liquid usedduring specimen-processing. Therefore, the ability to control hue viatemperature can be a useful feature. Temperature adjustment can also beused in conjunction with pH adjustment to achieve a desired hue, such asa desired level of bluing.

After stain setting and hue adjusting, the fourth washing can be used toremove residual stain-setting reagent from the specimens. The fourthwashing can include use of the same washing liquid discussed above inthe context of the first, second, and third washings. In at least somecases, the fourth washing includes a greater number of iterations thanthe first, second, and third washings, such as three instead of two.After the fourth washing, counterstaining the specimens can includedispensing a counterstaining reagent onto the slides, allowing thedispensed counterstaining reagent to remain in contact with thespecimens for a suitable counterstaining incubation time so as tocounterstaining the specimens (e.g., while the counterstaining reagentis in the form of a puddle having a shape maintained at least partiallyby surface tension), and then removing the dispensed counterstainingreagent. The counterstaining incubation time can be, for example, a timewithin a range from 30 seconds to 5 minutes. In a specimen-processingmethod in accordance with a particular embodiment of the presenttechnology, the counterstaining incubation time is 2 minutes.

The counterstaining reagent can be selected or formulated to adequatelycounterstain the specimens, such as to allow for proper differentiationbetween cytoplasmic and connective tissue. Furthermore, thecounterstaining reagent can be further selected or formulated to achievea desired stain hue, such as to have a pH that causes a desired stainhue. Counterstaining reagents selected or formulated in accordance withembodiments of the present technology can include deionized water, acounterstaining dye (e.g., eosin), and a solvent to maintain thecounterstaining dye in solution. As with the washing liquid, thestaining reagent, the stain-differentiating liquid, and thestain-setting reagent, the solvent can be a polyol, such as ethyleneglycol, propylene glycol, or a combination thereof. For example, thecounterstaining reagent can include greater than 30% by volume polyol,such as from 30% to 70% by volume polyol and, in some cases, from 40% to60% polyol. A counterstaining reagent selected or formulated inaccordance with a particular embodiment of the present technologyincludes about 500 mL deioni zed water, 750 milligrams of eosin Y, 1 mLof glacial acetic acid, and 500 mL of propylene glycol. Thecounterstaining reagent can have a pH, for example, within a range from3.65 to 4.25. This pH may be lower than the pH of conventional eosincounterstaining reagents. It may be possible, for example, to preventeosin Y from converting into a free acid at lower pH values (e.g., pHvalues less than 4) in propylene glycol than in ethanol. Counterstainingreagents selected or formulated in accordance with other embodiments ofthe present technology can include higher concentrations of eosin, suchas a concentration of 5.4 grams of eosin Y per liter. Thesecounterstaining reagents, for example, can rely heavily on regression toachieve a desired counterstain intensity.

After counterstaining, the fifth washing can be used to remove residualcounterstaining reagent from the specimens. The fifth washing can alsobe used to differentiate and regress the counterstain. When thecounterstain is an eosin counterstain, the counterstain differentiatingcan cause erythrocytes, collagen, and cytoplasm of muscle or epithelialcells within the specimens to be stained three different shades of pink,with cytoplasm having the lightest shade, erythrocytes having thedarkest shade, and collagen having an intermediate shade. Conventionalcounterstain differentiating and regressing is at least typicallycarried out in conjunction with dehydrating specimens. For example,conventional counterstain differentiating and regressing at leasttypically includes contacting specimens with graded ethanol and watermixtures having increasing concentrations of ethanol and decreasingconcentrations of water and then contacting the specimens with anhydrousalcohol.

The fifth washing can include use of the same washing liquid discussedabove in the context of the first, second, third, and fourth washings.In some cases, the duration of one or more iterations of the fifthwashing is adjustable to control the level of counterstaindifferentiating and regressing. For example, the fifth washing caninclude a first iteration during which the specimens are exposed to thewashing liquid for about 20 seconds, followed by a second iterationduring which the specimens are exposed to the washing liquid for aperiod of time within a range from 30 to 80 seconds. In aspecimen-processing method in accordance with a particular embodiment ofthe present technology, the period of time during which the specimensare exposed to the washing liquid during the second iteration is 50seconds. The first iteration can function primarily to remove residualcounterstaining reagent from the specimens. The second iteration canfunction primarily to allow for variable differentiating and regressingof the counterstain. Eosin staining tends to be relatively sensitive tounevenness associated with evaporation during counterstaindifferentiating and regressing. Thus, in at least some cases, the totaltime during which the specimens contact the washing liquid during thefifth washing is less than 100 seconds. The performance of the washingliquid for counterstain differentiating and regressing can influence itsformulation. For example, water concentrations significantly greaterthan 50% in the washing liquid may tend to cause non-standardcounterstain differentiating, such as cytoplasm of the specimens beingdarker than erythrocytes of the specimens. Water concentrationssignificantly less than 50% in the washing liquid may tend to produceinadequate levels of counterstain differentiating and regressing. Thus,as described above, the washing liquid can have a water concentration ofabout 50%, such as 50%+/−3%.

After the fifth washing, the specimens may have a residualhydrophilicity that would be incompatible with coverslipping. The secondconditioning of the specimens after the fifth washing can includereducing this hydrophilicity. In at least some cases, the secondconditioning includes dispensing a conditioning liquid onto the slides,allowing the dispensed conditioning liquid to remain in contact with thespecimens for a suitable period of time so as to wholly or incrementallycondition the specimens (e.g., while the conditioning liquid is in theform of a puddle having a shape maintained at least partially by surfacetension), and then removing the dispensed conditioning liquid. The timeduring which the dispensed conditioning liquid is in contact with thespecimens can be, for example, a time within a range from 5 seconds to15 seconds. In a particular example, the time is 10 seconds. Theconditioning liquid can be the same conditioning liquid used during thefirst conditioning. In at least some cases, in addition to being wellsuited for changing the hydrophobicity/hydrophilicity of specimens, theconditioning liquid is well suited for protecting specimens during thetime period between the fifth washing and coverslipping. For example,di(propylene glycol) ethers and tri(propylene glycol) ethers (e.g.,tri(propylene glycol) butyl ether) and other conditioning liquidsselected or formulated in accordance with embodiments of the presenttechnology may be superior to xylene for preventing potentiallydestructive drying of tissue during this time period. Thus, use of theseconditioning liquids may reduce or eliminate restrictions on the lengthof this time period. This can be useful, for example, to reduce timeconstraints on lockstep process management and/or to provide a timewindow during which additional operations can be performed on thespecimens.

As discussed above, conventional conditioning of specimens forcoverslipping is at least typically carried out in conjunction withcounterstain differentiating using graded ethanol and water mixturesfollowed by anhydrous ethanol. Thereafter, the specimens are at leasttypically contacted with xylene to stop the counterstain differentiatingand to further condition the specimens for interaction with acoverslipping adhesive. As discussed above in the context of the firstconditioning, however, use of ethanol and xylene in automatedhistological systems can be problematic, particularly when the systemsoperate at elevated baseline temperatures. Di(propylene glycol) etherand other conditioning liquids selected or formulated in accordance withembodiments of the present technology may reduce or eliminate the needfor ethanol. In at least some cases, the conditioning liquids partiallycondition the specimens for coverslipping and a coverslipping liquid isused in place of xylene after the conditioning liquid during the secondconditioning to further condition the specimens for interaction with acoverslipping adhesive. The coverslipping liquid can be selected orformulated to be immiscible with water (e.g., to reduce or eliminateleaching of dye from archived specimens) and to be volatile enough toadequately cure during a drying process of reasonable duration (e.g., 5minutes).

The coverslipping liquid can include a terpene, such as a monoterpene(e.g., limonene). A coverslipping liquid selected or formulated inaccordance with a particular embodiment of the present technologyincludes about 100% d-limonene with a suitable preservative, such as 500parts per million butylated hydroxytoluene. Use of monoterpenes in thecoverslipping liquid tends to be significantly less problematic than useof monoterpenes in the conditioning liquid. For example, the amount ofmonoterpene coverslipping liquid sufficient to prepare specimens forcoverslipping following use of di(propylene glycol) ether conditioningliquid can be far less than the amount of the di(propylene glycol) etherconditioning liquid used during the first conditioning and the initialphase of the second conditioning. In at least some cases, the utilizedamount of monoterpene coverslipping liquid is low enough that it fullyevaporates after its use without causing noticeable noxious fumes. Inthese cases, since there may be no liquid monoterpene waste, there mayalso be no need for special protocols, if any, for remediating and/orhandling of system waste liquids due to the presence of monoterpenes inthese liquids.

In automated histological systems configured in accordance withembodiments of the present technology, the coverslipping liquid can beapplied to specimens within a stainer, within a coverslipper after thespecimens exit the stainer, or at another suitable location. Use of thecoverslipping liquid can include first dispensing the coverslippingliquid onto the slides and then removing the dispensed coverslippingliquid. For example, the coverslipping liquid can be dispensed near theedges of the slides and swept across the slides using an air knife. Thiscan serve to remove any residual conditioning liquid remaining on theslides. Thereafter, the coverslipping liquid can be dispensed once,twice, three times, or another suitable number of times near the centersof the slides and left in place while coverslips are applied to theslides.

As discussed above, staining reagents and counterstaining reagentsselected or formulated in accordance with embodiments of the presenttechnology can include non-ethanol solvents to respectively maintain thestain and counterstain in solution. It can be advantageous for thesesolvents to be common, such as the same, within the same chemical class,or otherwise functionally analogous. Furthermore, it can be advantageousfor one or more other liquids used in conjunction with a given stainingreagent and counterstaining reagent to include a solvent the same as,within the same chemical class as, or otherwise functionally analogousto the common solvent of the staining reagent and the counterstainingreagent. This use of a common solvent is expected to enhancespecimen-processing consistency and quality. This benefit, for example,may be associated with enhanced efficiency and/or consistency with whicha given liquid displaces residual amounts of a previously dispensedliquid when the liquids have a common solvent. Other supplemental oralternative benefits and mechanisms are also possible.

In sets of liquids selected or formulated in accordance with at leastsome embodiments of the present technology, a staining reagent, acounterstaining reagent, and a washing liquid individually includegreater than 10% by volume polyol. In at least some of these and othersets of liquids selected or formulated in accordance with embodiments ofthe present technology, all, all but one, or all but two of a stainingreagent, a stain-differentiating liquid, a stain-setting reagent, acounterstaining reagent, and a washing liquid include greater than 10%by volume polyol, such as greater than 10% by volume of the same polyol,such as greater than 10% by volume propylene glycol. Inspecimen-processing methods in accordance with at least some embodimentsof the present technology, a total of all liquid dispensed onto slidesafter the slides are moved into a stainer (e.g., into atemperature-controlled internal environment of a stainer) and before theslides exit the stainer has a greater volumetric concentration of polyolthan of monohydric alcohol. In at least some cases, the total liquiddispensed is at least substantially free of monohydric alcohol or atleast has a volumetric concentration of monohydric alcohol less than 3%.Furthermore, the total liquid dispensed can be at least substantiallyfree of xylene.

Due, at least in part, to use of relatively few (e.g., one) conditioningliquid formulations, use of the same liquid for both washing andcounterstain differentiating, the ability to achieve a full range ofstaining intensities with relatively few (e.g., one) staining reagentformulation, and/or other factors, specimen-processing methods inaccordance with embodiments of the present technology can include use offewer different types of liquids than would be used during conventionalspecimen-processing methods. Similarly, complete sets of liquidsselected or formulated in accordance with embodiments of the presenttechnology can include fewer constituent liquids than conventional setswith corresponding functionality. Liquids belonging to sets of liquidsselected or formulated in accordance with embodiments of the presenttechnology can be respectively held in and drawn from differentcorresponding supply containers of automated histological systemsconfigured in accordance with embodiments of the present technology.These systems can be fluidically self-contained and operable with fewersupply containers, plumbing lines, and/or other liquid-handlingcomponents than are included in conventional systems of correspondingfunctionality. Among other potential benefits, this can reduce the cost,complexity, and/or bulk of automated histological systems configured inaccordance with at least some embodiments of the present technology.

The selection of processing liquids, the order in which the selectedprocessing liquids are dispensed, the number of dispensing and removingiterations for each processing liquid, and the duration ofliquid-to-specimen contact (e.g., incubation time) for each iterationcan be based on a predetermined recipe. In at least some cases,specimens immersed in a given liquid volume are at least partiallyuncovered before being contacted with another liquid volume of the sameprocessing liquid (e.g., in a subsequent iteration of the sameprocessing operation) or of a different processing liquid (e.g., tobegin a new processing operation). As discussed above, this may enhancethe performance (e.g., precision) of at least some specimen-processingoperations. In some cases, these enhancements are more pronounced in thecontext of progressive staining than in the context of regressivestaining. As such, there may be less need for stain differentiating andregressing in at least some specimen-processing methods in accordancewith embodiments of the present technology than there is in conventionalspecimen-processing methods.

Specimen-processing methods in accordance with embodiments of thepresent technology can include, within a stainer, automaticallydispensing liquids of no more than 6 different formulations onto slidesaccording to a predetermined recipe for at least deparaffinizing,staining, stain setting, counterstaining, and counterstaindifferentiating specimens carried by the slides. A complete set ofliquids for executing a methods can include a deparaffinizing liquid, aconditioning liquid, a staining reagent, a stain-setting reagent, acounterstaining reagent, and a washing liquid. Similarly,specimen-processing methods in accordance with embodiments of thepresent technology can include, within a stainer, automaticallydispensing liquids of no more than 7 different formulations onto slidesaccording to a predetermined recipe for at least deparaffinizing,staining, stain differentiating, counterstaining, and counterstaindifferentiating specimens carried by the slides. A complete set ofliquids for executing these methods can include a deparaffinizingliquid, a conditioning liquid, a staining reagent, astain-differentiating liquid, a stain-setting reagent, a counterstainingreagent, and a washing liquid. Other liquids that can be included inthese and other sets of liquids selected or formulated in accordancewith embodiments of the present technology include, for example, acoverslipping liquid and a cleaning liquid. In at least some cases, allconstituents of complete sets of liquids selected or formulated inaccordance with embodiments of the present technology are configured foruse without dilution.

Selected Examples of Support Systems

FIG. 89 is a perspective view of the liquid supply 6100 in accordancewith an embodiment of the technology. The liquid supply 6100 can includeone or more pumps 6110, filters 6112 (one identified), and a containerbay 6120. The container bay 6120 can include a series of container slots6122 (one identified) for holding containers. Containers holdingprocessing liquids can be placed in the slots 6122 and connected to thevarious pumps 6110, which pump the processing liquids to the stainers 6.FIG. 89 shows a container 6130 positioned in a slot 6122 and anothercontainer 6132 ready to be inserted into another slot 6122. When acontainer is empty, the liquid supply 6100 can automatically switch overto another container and, in some embodiments, can alert a user so thatthe empty container can be replaced with a new container withoutinterrupting system workflow. Processing liquids used in highquantities, such as deparaffini zing liquid and washing liquid, can besupplied from a bulk liquid container or multiple containers. A widerange of different fittings can be used to fluidically couple thecontainers to fluidic components of the liquid supply 6100.

The container 6132 can include one or more features for ensuring thatcorrect liquids are pumped into the appropriate components. The bay 6120can include one or more readers positioned to obtain processing-liquidinformation from each container, and such processing-liquid informationcan be part of a bar code, a magnetic element (e.g., a magnetic strip),or RFID tag. Where an RFID tag is included on the container 6132, thebay 6120 can read the RFID tag to confirm that the proper liquid hasbeen installed in the appropriate bay. Referring to FIGS. 2 and 89, thecontroller 18 (FIG. 2) can receive the information from the bay 6120 to(1) determine staining protocols based on available processing liquids,(2) track processing-liquid usage to determine scheduled containerreplacement, and/or (3) otherwise command components of the system 2based, at least in part, on the number and types of available processingliquids.

FIG. 90 is an isometric exploded view of the container 6132 inaccordance with an embodiment of the present technology. The container6132 can include a hat assembly 6200 and a receptacle 6202. The hatassembly 6200 can include arms 6210 for securely holding onto thereceptacle 6202 when arcuate members 6220 (one identified) of the arms6210 are positioned in a receiving feature 6230 (e.g., a through-holes,recesses, etc) of the receptacle 6202. The arms 6210 can be biasedinward to keep the arcuate members 6220 locked into the receivingfeature 6230. A user can pull the arms 6210 apart until the arcuatemembers 6220 are moved out of the receiving feature 6230 and can thenmove the hat assembly 6200 away from the receptacle 6202.

FIG. 91 is a partial cross-sectional view of the container 6132. The hatassembly 6200 and receptacle 6202 can have mateable handles 6300, 6302,respectively. When assembled, a user can conveniently grip the handles6300, 6302 to manually transport the container 6132. Other types ofhandle arrangements can also be used, if needed or desired. The hatassembly 6200 can include a conduit 6250 (e.g., a tubular member)extending downwardly through a chamber 6252 of the receptacle 6202. Anend 6254 of the conduit 6250 can be positioned at least proximate to abottom 6256 of the chamber 6252, or at any other desired location withthe chamber 6252. In some embodiments, the end 6254 can be positionedwithin at threshold distance (e.g., 0.5 inch (1.3 cm)) of the bottom6256. The conduit 6250 can have an angled section 6261 such that the end6254 is located adjacent to a side wall 6260 and positioned at thedeepest region of the chamber 6252 used to limit dead volume. The liquidcan be drawn through the conduit 6250 even when a minimal volume ofliquid is held by the receptacle 6202. As shown in FIG. 91, a relativelydeep region of the chamber 6252 can be positioned proximate to a sidewall 6260 of the receptacle 6202 to further minimize dead volumes, ifany.

The systems disclosed herein can also use other types of containers,including bag-in-the-box containers that include, without limitation,collapsible bags, tubes sealed into the bags, cover, and boxes.Non-exemplary embodiments of bag-in-the-box containers are disclosed inU.S. Pat. No. 7,303,725.

FIG. 92 is an isometric view of a waste container in accordance with oneembodiment. The waste container 7100 can include one or more sensorassemblies 7110 capable sensing the amount of liquid waste in a chamber7111. Waste can be delivered through feed tubes 7113 into the chamber7111. The sensor assemblies 7110 can include sensors 7115 and guide rods7120 along which sensors 7115 move in the vertical direction. The wastecontainer 7100 can be part of waste containers (e.g., waste containers32, 34 of FIG. 2) or at any other location within the system 2.

FIG. 93 is a cross-sectional view of the sensor 7115 in accordance withone embodiment of the present technology. The sensor 7115 can float tosense the volume of waste held in the chamber 7111 and can include afloat sensor 7142 and a protective shield 7144. The protective shield7144 can keep particles (e.g., precipitate from staining reagent) fromentering an sensor chamber 7145. The sensor 7142 and the protectiveshield 7144 can slide together along the rod 7120 while the protectiveshield 7144 prevents or limits substances (e.g., particles that canaffect operation of the sensor 7142) from entering the chamber 7145.Other configurations of sensors can be utilized.

CONCLUSION

This disclosure is not intended to be exhaustive or to limit the presenttechnology to the precise forms disclosed herein. Although specificembodiments are disclosed herein for illustrative purposes, variousequivalent modifications are possible without deviating from the presenttechnology, as those of ordinary skill in the relevant art willrecognize. In some cases, well-known structures and functions have notbeen shown or described in detail to avoid unnecessarily obscuring thedescription of the embodiments of the present technology. Although stepsof methods may be presented herein in a particular order, in alternativeembodiments the steps may have another suitable order. Similarly,certain aspects of the present technology disclosed in the context ofparticular embodiments can be combined or eliminated in otherembodiments. Furthermore, while advantages associated with certainembodiments may have been disclosed in the context of those embodiments,other embodiments can also exhibit such advantages, and not allembodiments need necessarily exhibit such advantages or other advantagesdisclosed herein to fall within the scope of the present technology. Forexample, while processing liquids selected or formulated in accordancewith some embodiments of the present technology are free of monohydricalcohol (e.g., ethanol) and/or xylene, processing liquids selected orformulated in accordance with other embodiments of the presenttechnology may include monohydric alcohol (e.g., ethanol) and/or xylene.This disclosure and associated technology can encompass a variety ofembodiments not expressly shown or described herein.

Certain aspects of the present technology may take the form ofcomputer-executable instructions, including routines executed by acontroller or other data processor. In at least some embodiments, acontroller or other data processor is specifically programmed,configured, and/or constructed to perform one or more of thesecomputer-executable instructions. Furthermore, some aspects of thepresent technology may take the form of data (e.g., non-transitory data)stored or distributed on computer-readable media, including magnetic oroptically readable and/or removable computer discs as well as mediadistributed electronically over networks. Accordingly, data structuresand transmissions of data particular to aspects of the presenttechnology are encompassed within the scope of the present technology.The present technology also encompasses methods of both programmingcomputer-readable media to perform particular steps and executing thesteps.

The methods disclosed herein include and encompass, in addition tomethods of practicing the present technology (e.g., methods of makingand using the disclosed devices and systems), methods of instructingothers to practice the present technology.

For example, a method in accordance with a particular embodimentincludes positioning a slide carrier at a first position while the slidecarrier holds a plurality microscope slides, robotically moving theslide carrier from the first position to a second position to move theslide carrier into a circulation loop defined by a heater apparatus, andconvectively heating the slides while the slide carrier is at the secondposition. A method in accordance with another embodiment includesinstructing such a method.

Throughout this disclosure, the singular terms “a,” “an,” and “the”include plural referents unless the context clearly indicates otherwise.Similarly, unless the word “or” is expressly limited to mean only asingle item exclusive from the other items in reference to a list of twoor more items, then the use of “or” in such a list is to be interpretedas including (a) any single item in the list, (b) all of the items inthe list, or (c) any combination of the items in the list. Additionally,the terms “comprising” and the like are used throughout this disclosureto mean including at least the recited feature(s) such that any greaternumber of the same feature(s) and/or one or more additional types offeatures are not precluded. Directional terms, such as “upper,” “lower,”“front,” “back,” “vertical,” and “horizontal,” may be used herein toexpress and clarify the relationship between various elements. It shouldbe understood that such terms do not denote absolute orientation.Reference herein to “one embodiment,” “an embodiment,” or similarformulations means that a particular feature, structure, operation, orcharacteristic described in connection with the embodiment can beincluded in at least one embodiment of the present technology. Thus, theappearances of such phrases or formulations herein are not necessarilyall referring to the same embodiment. Furthermore, various particularfeatures, structures, operations, or characteristics may be combined inany suitable manner in one or more embodiments.

1.-15. (canceled)
 16. An apparatus for heating a plurality of specimenscarried by a plurality of microscope slides, the apparatus comprising: ahousing at least partially defining a circulation loop; a blowerpositioned to produce a fluid flow along the circulation loop; a doorassembly moveable between a first position and a second position,wherein— when in the first position, the door assembly is configured toreceive a slide carrier that carries the plurality of microscope slides,and when in the second position, the door assembly is configured to holdthe slide carrier at a vertically-oriented position along thecirculation loop; and a heat source configured to heat the fluid flowalong the circulation loop such that the specimens are convectivelyheated by the fluid flow when the door assembly holds the slide carrieralong the circulation loop.
 17. The apparatus of claim 16, wherein thesecond position is angled relative to a horizontal plane.
 18. Theapparatus of claim 16, wherein the second position is substantiallyvertical relative to a horizontal plane.
 19. The apparatus of claim 16,further comprising a turbulence promoter positioned along thecirculation loop and configured to create a turbulent fluid flow along aturbulent flow portion of the circulation loop.
 20. The apparatus ofclaim 16, further comprising a laminar flow promoter positioneddownstream of the first portion of the circulation loop and configuredto create a laminar fluid flow along a laminar flow portion of thecirculation loop.
 21. The apparatus of claim 16, further comprising: afirst flow modifier positioned along the circulation loop and configuredto promote a turbulent fluid flow along a first portion of thecirculation loop; and a second flow modifier positioned downstream ofthe first portion of the circulation loop and configured to promote alaminar fluid flow along a second portion of the circulation loop. 22.An apparatus for heating a plurality of specimens carried by a pluralityof microscope slides, the apparatus comprising: a housing at leastpartially defining a circulation loop; a blower positioned to producefluid flow along the circulation loop; a first flow modifier positionedalong the circulation loop and configured to create a turbulent fluidflow along a first portion of the circulation loop; a second flowmodifier positioned downstream of the first portion of the circulationloop and configured to create a laminar fluid flow along a secondportion of the circulation loop; a door assembly moveable between anopen position and a closed position, wherein— when in the open position,the door assembly is configured to receive a slide carrier that carriesthe plurality of microscope slides, and when in the closed position, thedoor assembly is configured to position the slide carrier along thesecond portion of the circulation loop; and a heat source configured toheat the fluid flow such that the specimens are convectively heated bythe fluid flow when the door assembly holds the slide carrier along thesecond portion of the circulation loop.
 23. The apparatus of claim 22,wherein the door assembly holds the slide carrier such that theplurality of microscope slides are at an angle of inclination between 70degrees and 90 degrees relative to a horizontal plane when the doorassembly is in the closed position.
 24. The apparatus of claim 22,wherein the first flow modifier is a perforated plate.
 25. The apparatusof claim 22, wherein the second flow modifier includes one or morecurved vanes.
 26. The apparatus of claim 22, wherein the second flowmodifier comprises one or more tapered flow channels.
 27. The apparatusof claim 22, wherein, when the door assembly is in the closed position,the door assembly is positioned to hold the slide carrier such that afirst row of the slides held by the carrier is positioned above a secondrow of slides held by the slide carrier.
 28. The apparatus of claim 22,wherein, when the door assembly is in the closed position, the doorassembly is positioned to hold the slide carrier such that a first rowof the slides held by the carrier is positioned above and horizontallyspaced apart from a second row of slides held by the slide carrier. 29.A method for curing a plurality of specimens, wherein each of theplurality of specimens is covered by a coverslip and carried by one of aplurality of microscope slides, the method comprising: positioning aslide carrier at a first position while the slide carrier holds theplurality of microscope slides; robotically positioning the slidecarrier at a second position within a circulation loop defined by aheater apparatus; and convectively heating the coverslips and microscopeslides within the circulation loop while the slide carrier is at thesecond position.
 30. The method of claim 29, wherein convectivelyheating the coverslips and microscope slides includes providing a heatedfluid flow along a longitudinal length of one or more of the slides. 31.The method of claim 30, wherein the heated fluid flow flows across theslides at an average flow rate between about 5 m/s and 7 m/s.
 32. Themethod of claim 29, wherein convectively heating the coverslips andmicroscope slides includes convectively heating the coverslips and/orslides to a temperature in a range of about 90 degrees to about 100degrees.
 33. The method of claim 29, wherein convectively heating thecoverslips and microscope slides includes convectively heating thecoverslips and/or the microscope slides while the microscope slides arepositioned substantially horizontally.
 34. The method of claim 29,further comprising curing convectively the coverslips onto respectiveslides.